Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi User Guide

June 9, 2024
Thermo Fisher SCIENTIFIC

Invitrogen
Qubit™ 4 Fluorometer
Catalog Number Q33226
Publication Number MAN0017209
Revision D.0

MAN0017209 Qubit 4 Fluorometer With WiFi

For Research Use Only. Not for use in diagnostic procedures.

Manufacturer: Life Technologies Holdings Pte Ltd | Block 33 | Marsiling Industrial Estate Road 3 | #07-06, Singapore 739256 Information in this document is subject to change without notice.
DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
Revision history: MAN0017209

Revision Date Description
D.0 20 October 2021 Adding various edits, including deleting “Register your

instrument” topic and product edits in “Ordering Information”.
C.0| 09 April 2018| Add note about the requirement for driver installation in “Export data directly to computer”.
B.0| 30 January 2018| Update Safety and EMC standards and Results screen images for quality assays.
A.0| 05 October 2017| New user guide.

Important Licensing Information: This product may be covered by one or more Limited Use Label Licenses. By use of this product, you accept the terms and conditions of all applicable Limited Use Label Licenses.
Trademarks: All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Windows is a registered trademark of Microsoft Corporation.

© 2021 Thermo Fisher Scientific Inc. All rights reserved.

About this guide

Overview
This user guide describes how to operate the Qubit™ 4 Fluorometer.

User documentation
The guides listed below are available with the Qubit™ 4 Fluorometer.

Guide Pub. No.
Qubit™ 4 Fluorometer User Guide MAN0017209
Qubit™ 4 Fluorometer Quick Reference Card (QRC) MAN0017210

Additional resources are available on the Qubit™ Technical Resources page. Go to thermofisher.com/qubit to access protocols, application notes, and tutorials.

Text and keyboard conventions
Text and keyboard conventions used in the Qubit™ 4 Fluorometer User Guide are listed below. For safety alert words and symbols used in Thermo Fisher Scientific user documentation, see page 4.

Convention Use
Bold Bold text indicates user action. For example:

Click Run.
| Right arrow symbol ( ) indicates a menu choice and separates successive commands you execute or select from a drop-down or shortcut menu. For example: Select Settings ****Instrument settings.

User attention words

Two user attention words appear in Thermo Fisher Scientific user documentation.
Each word implies a particular level of observation or action as described below.

Note: Provides information that may be of interest or help but is not critical to the use of the product.
IMPORTANT! Provides information that is necessary for proper instrument operation, accurate installation, or safe use of a chemical.

Safety alert words
Four safety alert words appear in Thermo Fisher Scientific user documentation at points in the document where you need to be aware of relevant hazards. Each alert word—IMPORTANT, CAUTION, WARNING, DANGER—implies a particular level of observation or action, as defined below:

IMPORTANT! – Provides information that is necessary for proper instrument operation, accurate installation, or safe use of a chemical.
CAUTION! – Indicates a potentially hazardous situation that, if not avoided, may result in minor or moderate injury. It may also be used to alert against unsafe practices.
WARNING! – Indicates a potentially hazardous situation that, if not avoided, could result in death or serious injury.
DANGER! – Indicates an imminently hazardous situation that, if not avoided, will result in death or serious injury. This signal word is to be limited to the most extreme situations.

Except for IMPORTANT! safety alerts, each safety alert word in a Thermo Fisher Scientific document appears with an open triangle figure that contains a hazard symbol. These hazard symbols are identical to the hazard symbols that are affixed to Thermo Fisher Scientific instruments (see “Safety symbols” in Appendix C).

SDSs
The Safety Data Sheets (SDSs) for any chemicals supplied by Thermo Fisher Scientific are available to you free 24 hours a day. For instructions on obtaining SDSs, see “Safety Data Sheets (SDS)”.
IMPORTANT! For the SDSs of chemicals not distributed by Thermo Fisher Scientific contact the chemical manufacturer.

Product information

Product contents
The Qubit™ 4 Fluorometer is shipped with the components listed below:

Component Quantity
Qubit” 4 Fluorometer 1 each
Universal Power Cord with 4 adaptor plugs (for U.S. /E.U. /U.K. /Australia) 1

each
USB drive (includes the user guide)| 1 each
Qubit” 4 USB cable| 1 each
Qubit” 4 Fluorometer Quick Reference Card (QRC)| 1 each
Certificate of Conformity (COC)| 1 each
QubitT” screen cleaning cloth| 1 each

The complete user guide is also available for download at thermofisher.com/qubit.
See page 6 for the description and specifications of the Qubit™ 4 Fluorometer.

Upon receiving the instrument
Examine the instrument carefully for damage incurred during transit. Ensure that all parts of the instrument, including the accessories listed above, are included with the product. Damage claims must be filed with the carrier; the warranty does not cover in-transit damage.
See page 9 for instructions to set up the Qubit™ 4 Fluorometer.

Product description
Qubit™ 4 Fluorometer
The Qubit™ 4 Fluorometer is a benchtop fluorometer that can be used for the quantitation of DNA, RNA, microRNA, and protein, as well as for the measurement of RNA integrity and quality using the highly sensitive and accurate fluorescencebased Qubit™ assays. Additionally, Ion Sphere™ Particle quality can be evaluated on the Qubit™ 4  Fluorometer using the Ion Sphere™ Quality Control Kit prior to performing a sequencing run on the Ion Personal Genome Machine™ (PGM™) Sequencer. You can also use the Qubit™ 4 Fluorometer to directly measure the fluorescence of samples or to create new assays using the MyQubit software preprogrammed into the instrument.

Features

  • Fast and highly accurate quantitation of DNA, RNA, and protein in less than 3 seconds per sample.
  • Measure percentage of intact and degraded RNA in the sample in less than 5 seconds per sample.
  • High levels of accuracy using only 1–20 µL of sample, even with very dilute samples.
  • Use of dyes selective for dsDNA, RNA, or protein minimizes the effects of contaminants in the sample.
  • Stores results from up to 1000 samples.
  • Large, state-of-the-art color touchscreen for easy workflow navigation.
  • Graphically displays up to 20 data points for quantitation assays.
  • Instrument indicates samples that are in the extended range or out of range in the graph, as well as in the simple sample output screen.
  • Saves sample data as a CSV (comma separated value) or PDF file.
  • Exports data to a USB drive or directly to your computer via a USB cable.
  • On-board Reagent Calculator provides instructions to prepare Qubit™ working solution using your sample and standard inputs.
  • Allows easy definition and saving of assay preferences.
  • Instrument user interface can be personalized to show only the frequently used assays, to add new assays, including user-defined assays created with the MyQubit software and web tool, and to display in the language of your choice including English, French, Spanish, Italian, German, simplified Chinese and Japanese.

Instrument exterior components
Top view

Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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Back view

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  1. Touchscreen is the user interface containing the controls for all the functions needed and displays data from the assays.
  2. Sample chamber is used to load the assay tube containing the sample into the fluorometer for analysis.
  3. USB cable port (type mini-B) allows you to transfer your data directly to your computer using the USB cable supplied with the instrument or any other similar USB cable.
  4. USB drive ports (Type A) allow you to transfer and save data to your computer using the USB flash drive supplied with the instrument (or any other similar USB drive) for record keeping and printing purposes.
  5. Power inlet connects the Qubit™ 4 Fluorometer to an electrical outlet using the supplied power cord and the appropriate plug, based on the electrical outlet configuration in your country.

**Product specifications

**

Physical characteristics

Instrument type: Benchtop fluorometer
Instrument dimensions: 5.4 in (w) × 10 in (l) × 2.2 in (h)

(13.6 cm × 25 cm × 5.5 cm); rectangular shape
Weight:| 26.2 oz (743 g)
Operating power:| 100–240 VAC, 1.0 A/1.2 A
Frequency:| 50/60 Hz
Electrical input:| 12 VDC, 2.5 A
Installation site:| Indoor use only
Operating temperature:| 10–40°C
Operating humidity:| 20–80% (non-condensing)

Technical specifications

Dynamic range: 5 orders of magnitude
Processing time: ≤5 seconds/sample
Light sources: Blue LED (max ~470 nm)

Red LED (max ~635 nm)
Excitation filters:| Blue 430–495 nm
Red 600–645 nm
Emission filters:| Green 510–580 nm
Red 665–720 nm
Detectors:| Photodiodes; measurement capability from
300–1000 nm
Calibration type:| 2- or 3-point standard
Tube type:| 0.5-mL Real Time PCR (polypropylene) tubes
Warm-up time:| <35 seconds

USB drive

Capacity: 4 Gigabyte

Getting started

Set up the Qubit™ 4 Fluorometer
Set up the instrument

The Qubit™ 4 Fluorometer is a stand-alone instrument that does not require connection to a computer.

  1. After unpacking the instrument, place the instrument on a flat, level, dry surface.

  2. Plug one end of the supplied power cord into the Qubit™ 4 Fluorometer.

  3. Attach the appropriate plug adaptor to the other end of the power cord.

  4. Plug the power cord into the electrical outlet.
    Note: Ensure the plug of power adaptor remains accessible to allow disconnection.

  5. The instrument automatically powers on, first displaying the splash screen, and then the Home screen.

  6. To power down the Qubit™ 4 Fluorometer, unplug it.

IMPORTANT! Use the power cord plug adapter supplied with the instrument that is appropriate for the electrical outlet configuration in your country. Powering the instrument with an unapproved power cord may damage the instrument.

Home screen
When the instrument is powered on, the Home screen is automatically displayed. From the Home screen, you can:

  • Select one of the assays: dsDNA, RNA, oligo (ssDNA), protein, or Ion Sphere™.
  • Select the Fluorometer mode (the instrument behaves like a mini-fluorometer).
  • Access saved data.
  • Configure instrument settings.
  • Use the Reagent Calculator to determine the exact volumes of Qubit™ buffer and reagent required to prepare the Qubit™ working solution.

About Instrument screen

The About Instrument screen displays information about your Qubit™ 4 Fluorometer, including the currently installed software version. To access the About Instrument screen:

  1. On the Home screen, press Settings.

  2. On the Settings screen, press About Instrument to display the About Instrument screen.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  3. Press Done, Cancel, or Back ( ) to return to the Settings screen.

Optional: Configure instrument settings
Introduction
You can configure the following instrument settings for the Qubit™ 4 Fluorometer:

  • Sleep mode (page 12)
  • Brightness (page 12)
  • Date/Time (page 13)
  • Reset instrument (page 14)
  • Language (page 15)

Access the Instrument settingsscreen

  1. On the Home screen, press Settings.
  2. On the Settings screen, press Instrument settings to display the Instrument settings screen.

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Adjust the sleep mode

The Qubit™ 4 Fluorometer has a sleep mode (i.e., automatic standby) that is triggered by inactivity. The system default is 10 minutes of inactivity before the instrument goes into sleep mode. To adjust this time:

  1. On the Instrument settings screen (page 11), press Sleep mode to display the Sleep mode screen.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  2. Enter the time in minutes allowed before the instrument goes into sleep mode by pressing the minutes field, then using the number pad to select a value. The software requires a minimum of 1 minute and a maximum of 60 minutes.

  3. Press Done to save the changes and return to the Instrument settings screen. Press Cancel or Back ( ) to return to the Instrument settings screen without saving the changes.

Adjust screen brightness

  1. On the Instrument settings screen (page 11), press Brightness to display the Brightness screen.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  2. Move the slider button up or down to adjust the brightness of the display.

  3. Press Done to save the changes and return to the Instrument settings screen.
    Press Cancel or Back ( ) to return to the Instrument settings screen without saving the changes.

Adjust the date andtime
The date and time are preset when you receive the instrument. To adjust the date and time:

  1. On the Instrument settings screen (page 11), press Date/Time.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
8

  2. Select a date format, select a time format, then press Next.

  3. Enter the date and time:
    a. Press a date field (day, month, or year), then use the number pad to select a value.
    b. Press a time field (hours or minutes), then use the number pad to select a value.
    c. Select AM or PM.

  4. Press Done to save the changes and return to the Instrument settings screen.
    Press Cancel or Back ( ) to return to the Instrument settings screen without saving the changes.

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Reset function

The Reset function returns the Qubit™ 4 Fluorometer to its default factory settings, and erases all saved data, user–defined instrument settings, and custom assays.

IMPORTANT! The reset function is not reversible.

To return the instrument to its default factory settings:

  1. On the Instrument settings screen (page 11), press Reset instrument to display the Reset Qubit™ 4 screen.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  2. Press Reset. After the reset is complete, the Home screen is displayed. All data, user–defined instrument settings, and custom assays are removed.
    Press Cancel or Exit ( ) to return to the Instrument settings screen without saving the changes.

Change the display language

You can change the language that the Qubit™ 4 Fluorometer displays to English (default), French, German, Italian, Spanish, simplified Chinese, and Japanese.

  1. On the Instrument settings screen (page 11), press Language to display the Language screen.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
11

  2. Select a language, then press Next.

  3. Press Yes to confirm the change and return to the Instrument settings screen.
    Press Cancel or Exit ( ) to return to the Instrument settings screen without saving the changes.

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Guidelines for using the Qubit™ 4 Fluorometer

Recommendations
To obtain the best results, follow the recommendations below. For more information, see “Critical Qubit™ Assay considerations”, page 59.

  • Do not operate the instrument in direct sunlight.

  • Wear gloves during sample handling.

  • Use the instrument at room temperature only (22–28ºC).

  • Bring all kit reagents to room temperature and insert all assay tubes into the instrument only for as much time as it takes for the instrument to measure the fluorescence.

  • Do not hold the assay tubes in your hand before performing a measurement.

  • Make sure that you have calibrated the Qubit™ 4 Fluorometer using the appropriate standards.

  • Incubate the tubes for the Qubit™ DNA and RNA assays for 2 minutes after mixing the sample or standard with the working solution.

  • Incubate the tubes for the Qubit™ protein assays for 15 minutes after mixing the sample or standard with the working solution.

  • Incubate the tubes for the Qubit™ Protein Broad Range Assay for 10 minutes after mixing the sample or standard with the Protein BR Assay Reagent.

  • If you are performing multiple readings of a single tube, remove the tube from the instrument and let it equilibrate to room temperature for 30 seconds before taking another reading.
    Note: Multiple readings of RNA samples is not recommended.

  • Visit thermofisher.com/qubit for additional application notes, technical notes, citations, software updates, and a list of validated Qubit™ assays that have been tested using the Qubit™ 4 Fluorometer.

Assay tubes for the Qubit™ 4 Fluorometer

Only thin-wall, clear 0.5-mL PCR tubes are appropriate for use in the Qubit™ 4 Fluorometer. Acceptable tubes include Qubit™ assay tubes (Cat. No. Q32856, 500 tubes). The minimum assay volume must be 200 µL for an accurate read.

Perform assays

Overview
This section provides instructions for using the Reagent Calculator, running new calibration standards or for using the last calibration, and for reading the samples using a Qubit™ assay kit.

Materials needed

  • A Qubit™ assay kit appropriate for quantifying your samples (see page 61 for available Qubit™ assay kits and ordering information)

  • DNA, RNA, or protein samples

  • Qubit™ assay tubes or other appropriate 0.5-mL assay tubes

  • Appropriate standards for the assay you are performing
    Note: For instructions on the preparation of these standards, see the instructions that accompany the assay you are using or the Qubit™ 4 Fluorometer Quick Reference Card (QRC) (Pub. No. MAN0017210).

  • (Optional) USB drive or USB cable for data transfer, supplied with the instrument or available separately

Optional: Use Reagent Calculator to prepare Qubit™ Working Solution

Introduction
Use the on-board Reagent Calculator to quickly determine the correct amount of Qubit™ dye and buffer required to make the appropriate amount of Qubit™ Working Solution required to prepare your samples and standards.
Note: The Reagent Calculator is not intended for use with the Qubit™ Protein Broad Range Assay.

Use the Reagent Calculator

  1. On the Home screen, press Reagent Calculator.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  2. Enter the total number of samples and standards that you will be running on the Qubit™ 4 Fluorometer.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  3. Optional: Select Include overage if you want to include an additional tube in the total calculated volume.

  4. Press Enter to calculate the amount of Qubit™ dye and buffer required to prepare the Qubit™ Working Solution with these inputs.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
16 Note: You can change the total number of tubes that you plan to run or the overage selection on this screen.

  5. Press Done to return to the Home screen to run your Qubit™ assay.

Calibrate assays on the Qubit™ 4 Fluorometer
Introduction
For each assay, you have the choice to run new standards for calibrating the assay on the Qubit™ 4 Fluorometer or to use the values from the previous calibration (for more information, see “Qubit™ 4 Fluorometer”, page 60).
Note: The Reagent Calculator is not intended for use with the Qubit™ Protein Broad Range Assay.

Read new standards for calibration

  1. On the Home screen, select the assay type for which you wish to read new standards.
    To go to the second page of available assays, swipe the screen to the right. To return to the first page, swipe the screen to the left.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  2. Select the desired assay.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
18 Note: The software displays the available assays for the assay type you selected in step 1. The example in step 2 shows the available assays for the dsDNA assay type.

  3. If you have already performed a calibration for the selected assay, the software prompts you tochoose between reading new standards and running samples using the previous calibration. Press Read standards.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
19 Note: To apply the previous calibration to your sample readings, press Run samples. The software prompts you to insert the assay tube containing sample. See “Read samples”, page 24.

  4. At the prompt, insert Standard #1 into the sample chamber and press Read standard. Be sure to use the Standard #1 appropriate for the assay you are performing. The reading takes approximately 3 seconds.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  5. At the prompt, insert Standard #2 into the sample chamber and press Read standard. Be sure to use the Standard #2 appropriate for the assay you are performing.

  6. For Qubit™ protein and RNA IQ assays only: At the prompt, insert Standard #3 into the sample chamber and press Read standard.

  7. The calibration is complete after Standard #2 is read (or after Standard #3 is read for the Qubit™ protein or RNA IQ assay). The software displays the results.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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Calibration results

  • For Qubit™ quantitation assays: If the calibration is successful, the software displays the Read standard screen with the Fluorescence vs. Concentration graph.

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In the Fluorescence vs. Concentration graph, the standard data points are connected by a line and open circles represent correct standards.

  • For the RNA IQ assay: If the calibration is successful, the software displays the Ready for samples message.

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23

  • If the calibration is not successful, the software displays the Calibration error message (page 23).

Note: The Read standard screen displays the raw fluorescence values for Standard #1 and Standard #2 (and Standard #3, if applicable). These values can assist you in making a judgment regarding the calibration results. For the Qubit™ dsDNA BR, Qubit™ dsDNA HS, Qubit™ ssDNA, Qubit™  microRNA, Qubit™ RNA HS, Qubit™ RNA BR, Qubit™ RNA XR, and Qubit™ protein assays, the reading given by Standard #2 should be at least ten times higher than that of Standard #1. For the Qubit™ protein assay, the reading given by Standard #3 should be at least 40% higher than that of Standard #2. For the Qubit™ RNA IQ assay, the instrument verifies the calibration parameters at each stage of the three point calibration process, confirming the appropriate response of the assay to the standards provided in the kit. If any issues arise during calibration, the instrument will guide you to appropriate actions to correct the issue.

Calibration error
If you receive the “Calibration error” message, you can re-run the standards:

  1. In the Error screen, press OK.
  2. Review the Read standard screen (see Note on page 23).
  3. If you wish to re-run the standards, or run new standards, press Read standards, then repeat the calibration procedure (beginning on page 18).

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Note: For the RNA IQ assay, specific standards can be re-run depending on the calibration error. The error message provides guidance on how to correct the calibration error.

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Read samples
Before you begin

  • Calibrate the Qubit™ 4 Fluorometer as described on page 18. (Run the appropriate standards or accept the values from the previous calibration.)
  • Prepare the samples. Refer to the instructions provided with the assay.

Note: Incubate the samples for the appropriate amount of time after mixing them with the working solution (2 minutes for the Qubit™ DNA and RNA assays, 15 minutes for the Qubit™ protein assay, or 10 minutes for the Qubit™ Protein Broad Range Assay).

Read samples

  1. In the Read standard (for Qubit™ quantitation assays) or Ready for samples screen (for Qubit™ RNA IQ assay), press Run samples.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  2. In the Sample volume screen, select the sample volume and units for the Qubit™ quantitation assays or the Qubit™ RNA IQ assay:
    a. Press the + or – buttons on the wheel to select the sample volume added to the assay tube (between 1 and 20 µL).
    b. For the Qubit™ quantitation assays, select the units for the output sample concentration from the dropdown menu.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
27 Note: In the RNA IQ assay, there are no units associated with the RNA IQ value (i.e., RNA IQ#) and only sample volume is required.

  3. Insert a sample tube into the sample chamber, close the lid, and then press Read tube. The reading takes approximately 3 seconds for the quantitation assays and approximately 5 seconds for the quality assays.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  4. The software displays the results. See “Results screen” on page 26.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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Read multiple samples
To read multiple samples for the same assay:

  1. Remove the current sample, and insert a new sample.
  2. To change the sample volume, swipe right or press the left arrow.
  3. Press Read tube.
  4. Repeat step 2 (page 24) and step 3 (page 25).

Results
Results screen Sample quantitation

The Results screen displays the results of the sample run.

  • If the results are within the assay’s range, the concentration values are displayed.
    The top value (in large font) is the concentration of the original sample. The bottom value is the dilution concentration (the concentration of the sample in the tube inserted into the Qubit™ 4 fluorometer).

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  • If the results are outside of the assay’s range, an “Out of Range” message is displayed.

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31

  • To return to the sample volume screen, swipe right or press the left arrow.
  • To view the Fluorescence vs. Concentration graph, swipe left or press the right arrow. In the graph:
    -­ Open circles represent correct standards.
    -­ The large gray circle represents the most recent sample.
    -­ Blue circles represent samples that fall within the assay’s core range.
    -­ Yellow circles represent samples that fall within the assay’s extended range.
    -­ Red circles represent samples or standards that fall outside the assay’s range.

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33

  • To return to the previous screen, swipe right or press the left arrow.

Note: The Qubit™ 4 Fluorometer automatically stores numeric data from all reads, but does not store graphic data.

Results screen Sample quality

The Results screen displays the results of the sample run.

  • If the results are within the assay’s range, the sample quality values are displayed. The result presented in the circle (i.e., RNA IQ#) is the total value for the RNA sample integrity and quality, which is the calculated percentage of large and small RNA in the sample.

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The RNA IQ# is based on a scale of 1–10, similar to other RNA quality scores.
A small RNA IQ# in the assay indicates that the sample is comprised of mainly small RNA and a larger RNA IQ# indicates that the sample consists of mainly large RNA or structured RNA (mRNA, tRNA, rRNA).
The actual percentages of small or degraded RNA and large or intact RNA in the sample are displayed below the RNA IQ#.

Measurement error
If the results are outside of the assay’s range, a “Measurement error” message is displayed.

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Fluorometer mode
Introduction

You can use the Qubit™ 4 Fluorometer as a mini-fluorometer by selecting the Fluorometer mode.
The Fluorometer mode allows you to choose the excitation light source (blue LED or red LED) while reading fluorescence in both the green and far-red emission channels (for the blue LED) or in the far-red emission channel only (for the red LED).
The reading is in raw fluorescence units (RFU).

Read sample in Fluorometer mode

  1. On the Home screen, press Fluorometer.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  2. On the Choose excitation screen, select Blue (470 nm) or Red (635 nm).
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  3. Insert the sample, then press Read tube.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  4. For Blue excitation, the Qubit™ 4 Fluorometer displays both the green emission and the far red emission of your sample. For Red excitation, the Qubit™ 4 Fluorometer displays only the far red emission of your sample.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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Read multiple samples in Fluorometer mode

To read multiple samples in the fluorometer mode:

  1. Remove the current sample, and insert a new sample.
  2. Press Read tube.

Ion Sphere™ Assay

The Ion Sphere™ assay on the Qubit™ 4 Fluorometer enables quick and easy assessment of Ion Sphere™ Particles (ISPs) prior to performing a sequencing run on the Ion Personal Genome Machine™ (PGM™) Sequencer using the Ion Sphere™ Quality Control Kit.
All ISPs bind a complimentary, green-fluorescent Alexa Fluor™ 488 dye-labeled oligonucleotide, whereas only those beads that are template positive bind a complementary red-fluorescent Alexa Fluor™ 647 dye-labeled oligonucleotide. The Qubit™ 4 Fluorometer measures the fluorescence emission of each dye conjugate and a Microsoft Excel™ file template is used to calculate the percentage of template- positive ISPs from the ratio of the Alexa Fluor™ 647 dye signal to the Alexa Fluor™ 488 dye signal. This quality-control step using the Qubit™ Fluorometer determines the likelihood of a successful sequencing run, ultimately saving time and expense during subsequent sequencing.

Note: For more information about the Ion Sphere™ Quality Control Kit (Cat. No. 4468656), go to thermofisher.com and search for the catalog number.

Manage data

Introduction

The Qubit™ 4 Fluorometer can save data for up to 1000 samples.
For the saved data, the instrument allows you to:

  • View detailed data for each sample (page 32).
  • Rename data files (page 35).
  • Save data as a CSV (comma separated value) or PDF file, and export the file to a USB drive or directly to your computer (page 36).
  • Delete data files (page 40)

View detailed sample data

  1. From the Concentration, Graph, RNA IQ results, or Home screen, press Data.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
40

  2. On the Export data screen, press the data set of interest.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
41A Data screen displaying a list of data entries for that data set appears. To view additional data entries that do not fit in the screen, scroll up or down.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  3. To reverse the order of the data entries, press the blue chevron above the sample number column.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  4. To view the sample details, press the sample of interest. A Data details screen opens.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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Rename data files

  1. On the Concentration, Graph, RNA IQ results, or Home screen, press Data.

  2. On the Export data screen, select the data set of interest.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  3. From the list of data entries for the selected data set, press the sample of interest to open the Data details screen.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
46

  4. On the Data details screen, press the Sample # field (indicated by the red arrow), enter the new name using the keyboard that appears, then press Enter.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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Export data

Introduction

The Qubit™ 4 Fluorometer is designed for standalone use; it does not require an external computer. However, to archive data and generate reports, you can export the numeric data stored in the CSV or PDF file to a computer either by using the USB drive or by saving directly to the computer using the USB cable. You can then import the file into any spreadsheet program.

Export data using the USB drive

  1. Insert the USB drive into the Qubit™ 4 Fluorometer.

  2. On the Concentration, Graph, RNA IQ results, or Home screen, press Data.

  3. On the Export data screen, check the selection box to the left of each data set you wish to export.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
48

  4. To save only individual data entries from a data set, press the data set of interest, and then check the selection box to the left of the samples you wish to export.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
49

  5. Select CSV or PDF and press Export to export the data.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
50

  6. Transfer the USB drive to the USB drive port on your computer.

Export data directly to a computer

Data files may also be exported from the Qubit™ instrument to a computer using a USB cable such as the one provided with the Qubit™ instrument. The USB cable must remain connected to both the Qubit™ instrument and the computer during this process.
Note: The Windows® XP operating system requires the installation of a driver. The driver and installation instructions are available for download from Qubit Technical Resources at thermofisher.com/qubit.

To export data directly to a computer:

  1. Connect the Qubit™ instrument to the computer using the USB cable.
  2. From the computer, access the Qubit™
  3. Fluorometer as an external device.
  4. Access the files saved on the Qubit™ instrument using the operating system’s normal file browsing menus.
  5. Copy the desired files to the computer.

Note: Files cannot be exported directly to an external computer using the Export workflow on the Qubit™ instrument.

Information saved to the Export file

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Note: For the RNA IQ assay, there is no original sample concentration, Qubit™ tube concentration or dilution factor. Instead, the results file contains % large and % small RNA information.
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Export data to the cloud using Wi-Fi

Data from the Qubit™ instrument can also be exported to the cloud using a Wi- Fi connection.

Note: Confirm the Qubit™ 4 instrument has the latest firmware (under Settings

About Instrument) and correct date and time.
The lastest firmware is available at:
https://www.thermofisher.com/us/en/home/industrial/spectroscopyelemental- isotope-analysis/molecularspectroscopy/fluorometers/qubit/qubit-technical- resources.html.

  1. Insert the Wi-Fi dongle into the USB port, then power on the instrument.

  2. From the Home screen, press Settings > Instrument Settings, then select Network Connection.

  3. On the Choose Network screen, select the network to join.
    a. On the Configure & Join Network screen, enter the password into the PSK box.
    b. Press Join.
    The network will be highlighted in a brighter blue color once connected.

  4. On the Home screen, press Data.

  5. On the Data screen, check the selection box to the left of each data set you wish to export.
    a. To select all data sets, press the blue check icon in the header row.
    b. To export only individual data entries from a data set, press the data set of interest, then check the selection box to the left of the samples desired for export.

  6. After selecting the data sets or sample runs, press Actions, then Export.

  7. In the Export data screen, select the Export method.

  8. Press Export.

Export data to the cloud using your Connect account

  1. Register or sign in to your account at https://www.thermofisher.com/us/en/home/digital-science.html.

  2. Click the Instrument Connect icon on the left side toolbar, then click on the blue Add Instrument icon in the upper right corner.

  3. Select Qubit_4 from the drop-down menu and click Next. You are then instructed to “Insert linking code: Get the code from your instrument.”

  4. On the Qubit™ 4 instrument Home screen, click on the profile (person) icon on in the top left corner to go to the Sign In screen.

  5. Under New to this instrument?, select Get Started.

  6. Under Connect to the Thermo Fisher Cloud, select Connect.

  7. From the Connect to the Cloud screen on the Qubit™ 4 instrument, select Dashboard on PC.

  8. On the Home screen, press Data.

  9. On the Data screen, check the selection box to the left of each data set you wish to export.
    a. To select all data sets, press the blue check icon in the header row.
    b. To export only individual data entries from a data set, press the data set of interest, then check the selection box to the left of the samples desired for export.

  10. After selecting the data sets or sample runs, press Actions, then Export.

  11. In the Export data screen, select the Export method.

  12. Press Export.

Delete data
Delete data files

  1. On the Concentration, Graph, RNA IQ results, or Home screen, press Data.

  2. On the Export data screen, check the selection box to the left of each data set you wish to delete.
    To delete only individual data entries from a data set, press the data set of interest, and then check the selection box to the left of the samples you wish to delete.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  3. Press Delete. A warning screen appears.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
54

  4. Press Delete to permanently delete the sample data or data set.
    Press Cancel to return to the screen previously viewed without deleting any data.

Manage, import, and create assays

Overview
The Qubit™ 4 Fluorometer allows you to personalize the instrument’s user interface to hide and show (unhide) assays or folders and to move them to different locations. In addition, you can add custom MyQubit assays by importing them from our website at thermofisher.com/qubit or create new assays using the MyQubit software and web tool.

Manage assays
Manage assays function

The Manage assays function allows you to hide and show (unhide) assays or folders and to move them to different locations.

  1. To access the Manage assays function, press Settings on the Home screen. ****
  2. On the Settings screen, press Manage assays to open the Manage assays screen.
    From the Manage assays screen, you can select Show/hide assays (page 42) or Move assays (page 44).

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55

Show/Hide assays
Show/Hide all assays within the assay folder

  1. 1From the Settings screen, press Manage assays.

  2. On the Manage assays screen, press Show/hide assays.
    The software displays a list of the existing assay folders available on the instrument. Each assay folder contains individual assays.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
56

  3. Select the assay folder containing the assays you wish to show or hide.
    The software displays a list of existing individual assays within the selected assay folder and the controls for showing and hiding them.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
57

  4. To hide all of the assays in the assay folder, swipe the Assay folder On/Off switch located to the right of the assay folder name to the Off position (to the left).
    To show all of the assays in the assay folder, swipe the Assay folder On/Off switch to the On position (to the right).

  5. Press the Back button ( ) to save the changes and return to the previous screen, or press Done to save the changes and return to the Settings screen.

Show/Hide individual assays

  1. From the Settings screen, press Manage assays.

  2. On the Manage assays screen, press Show/hide assays.
    The software displays a list of the existing assay folders available on the instrument. Each assay folder contains individual assays.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
58

  3. Select the assay folder containing the assays you wish to show or hide.
    The software displays a list of existing individual assays within the selected assay folder and the controls for showing and hiding them.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
59

  4. To hide an individual assay, swipe the Assay On/Off switch located to the right of the assay to the Off position while keeping the Folder On/Off switch in the On position.
    To show an individual assay, swipe the respective Assay On/Off switch to the On position while keeping the Folder On/Off switch in the On position.

  5. Press the Back button ( ) to save the changes and return to the previous screen, or press Done to save the changes and return to the Settings screen.

Move assays
Move assay folders/ individual assays on the Home screen

  1. From the Settings screen, press Manage assays.

  2. On the Manage assays screen, press Move assays.
    The software displays the Manage assays screen with available assay folders and individual assays, and activates the move function.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
60

  3. Press and hold on the desired assay folder or individual assay to detach it from its current position, then drag it to its new location.
    To move the selected object to a different Home screen page, drag it up or down to the desired screen.

  4. Release the drag to drop the selected assay folder or individual assay in its new location.

  5. Press the Back button ( ) to save the changes and return to the Manage assays screen.

Move assays between folders

  1. From the Settings screen, press Manage assays.

  2. On the Manage assays screen, press Move assays.
    The software displays the Manage assays screen with available assay folders and individual assays, and activates the move function.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
61

  3. Press the desired assay folder to open the folder’s assay screen.

  4. Press and hold the individual assay to detach it and drag it over the Home screen icon ( ).

  5. Release the drag to drop the selected assay to the Home screen.

  6. Press the Home button ( ) to return to the Home screen.
    In the Home screen, you can press and drag the selected assay to another assay folder or location as described in “Move assay folders/ individual assays on the Home screen”, page 44.

  7. Press the Back button ( ) to save the changes and return to the Manage assays screen.

Move assays within folders

  1. From the Settings screen, press Manage assays.
  2. On the Manage assays screen, press Move assays.
  3. Press the desired assay folder to open the folder’s assay screen.
  4. Press and hold the individual assay to detach it, and drag and drop it to its new location in the folder.
  5. Press the Back button ( ) to save the changes and return to the Manage assays screen.

Import assays

MyQubit assay files
You can add pre-existing MyQubit assays to your Qubit™ 4 Fluorometer by downloading the assay files from thermofisher.com/qubit, then importing the files to the instrument from a USB drive or from an external computer via a USB cable.
Note: MyQubit assay files have the file extension .qbt.

Import assays from a USB drive

  1. To import one or more new assays, go to thermofisher.com/qubit and download the MyQubit assay files (.qbt file) you wish to install to the root irectory of a USB drive.
    Do not download the .qbt files to a folder in the USB drive.

  2. Insert the USB drive containing the new assay files into the USB drive port of the Qubit™ 4 Fluorometer.

  3. Go to the Settings screen and press Import new assay. The instrument searches the USB drive for assay files and the Import new assay screen displays the names of the assays found in the USB drive.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
62

  4. Press the assays you wish to import to select. Touching the selected assay a second time deselects it. You can select multiple assays for import.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  5. Press Next. The Import new assay screen displays the name of the assay being imported (or the first assay, if multiple assays were selected) and lists the destination folders where you can save it.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
64Press the Back button ( ) or Cancel to return to the Settings screen without importing the assays.
    Note: Each assay folder can contain up to 9 individual assays. The Home screen can contain up to 36 assays or folders.

  6. Select the destination folder for the new assay and press Save. The Verification screen opens (see Step 9, page 48).
    Alternatively, press New folder to create a new destination folder (see Step 8, page 48).
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
65Press the Back button ( ) or Cancel to abort the import process and return to the Settings screen.

  7. In the New folder screen, press the New folder text field. The keyboard opens.
    Using the keyboard, enter the name of the new destination folder. You can use up to 11 characters; spaces at the beginning or the end of the name are not recognized.
    Press Enter, then press Done to return to the Import new assay – Where screen.
    The newly created folder will appear on the list of folders.
    Select the new destination folder, then press Save. The Verification screen opens (see Step 9, page 49).
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  8. In the Verification screen, press Done to complete the import process.
    If importing only one assay, you will be returned to the Home screen.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
67If importing multiple assays, you will be returned to the Import new assay screen and prompted to select the destination folder for the next assay in your import queue (see Step 10, page 49).

  9. Select the destination folder for the next assay in your import queue, then press Save.
    Repeat this process until you have imported all the assays you have selected in Step 4 (page 46).
    Once all the assays you have selected are imported successfully, the Verification screen opens.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  10. Press Done to complete the import process and return to the Home screen.

Import assays from a computer

When the Qubit™ 4 Fluorometer is connected to a computer via a USB cable, MyQubit .qbt assay files can be transferred directly from the computer to the instrument.

  1. Go to thermofisher.com/qubit and download the MyQubit assay files (.qbt file) you wish to install.

  2. Connect the Qubit™ 4 Fluorometer from the USB cable port to the computer using a mini-B USB cable. The Qubit™ 4 Fluorometer should be recognized as an external device by the computer.

  3. Copy and paste or click-drag the .qbt files you wish to import to the Qubit™ instrument window on your computer.
    Note: The assays will not be visible on the Qubit™ 4 Fluorometer until you perform the next step.

  4. Go to the Settings screen on the Qubit™ 4 Fluorometer and press Import new assay. The Import new assay screen displays the names of the assays found on the computer.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  5. Follow Steps 4–11 of the import procedure described in “Import assays from a USB drive”, page 46.
    Note: The USB cable must remain connected to both the Qubit™ 4 Fluorometer and the computer for the assay to be imported.

Create your own assay for the Qubit™ 4 Fluorometer
Create a MyQubit assay

All Qubit™ 4 Fluorometers are pre-loaded with MyQubit software, which allows you to create new applications for the Qubit™ 4 Fluorometer using parameters uploaded to the instrument via a USB drive, without changing the existing assays. Fluorometer mode allows you to use the Qubit™ 4 Fluorometer with any reagent or assay compatible with the optical settings of the instrument. In this way, the device can be used as a mini-fluorometer. With the data gathered in the Fluorometer mode, personalized, custom assays can be created using the guided webtool and the MyQubit EZGuide available at thermofisher.com/qubit.

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Note: For complete instructions on creating your own assay and access to the web tool, go to thermofisher.com/qubit.

Instrument maintenance

Maintenance

The Qubit™ 4 Fluorometer does not need regular maintenance. To troubleshoot problems with the instrument, contact Technical Support (page 71).

  • Do not perform any repairs or service on the Qubit™ 4 Fluorometer to avoid damaging the instrument.
  • Do not expose the Qubit™ 4 Fluorometer to direct sunlight.

CAUTION! Never disassemble or service the instrument yourself. Do not remove any covers or parts that require the use of a tool. Unauthorized repairs may damage the instrument or alter its functionality, which may void your warranty. Contact your local distributor to arrange for service.

Clean the Qubit™ 4 Fluorometer

We recommend that you clean the Qubit™ 4 Fluorometer periodically to prevent the buildup of dust and dirt that might reduce its performance and cause contamination.

CAUTION! To avoid electrical shock, always disconnect the power cable before cleaning or decontaminating the instrument.
IMPORTANT! Using a cleaning or decontaminating method other than that specified by the manufacturer may result in damage to the instrument.

  • Clean the surface of the Qubit™ 4 Fluorometer with a damp cloth.
  • To clean the touchscreen, disconnect the power cable, and clean the touchscreen with a soft cloth lightly moistened with LCD (liquid crystal display) cleansing detergent.
  • Cleaning the screen with excessive force can damage the touchscreen. Wipe the screen dry immediately.
  • Do not use abrasive cleaning solutions or material to prevent the touchscreen from getting scratched.
  • To disinfect the instrument, disconnect the power cable from the Qubit™ 4 Fluorometer and clean the instrument, including the touchscreen, with a soft cloth lightly moistened with 70% ethanol, 70% isopropanol, or 10% bleach (0.6% sodium hypochlorite).
  • The cloth included with the instrument is not recommended for use with ethanol or isopropanol.
  • Ensure that the cleaning solution does not enter the power button, the power inlet, the sample port, or the USB drive ports.
  • Never pour or spray any liquids directly on the instrument to avoid electrical shock when the instrument is plugged in.

Software updates
Update the software

To update the Qubit™ 4 Fluorometer with the latest software from Thermo Fisher Scientific:

  1. Download the latest software to a USB drive or to your computer from thermofisher.com/qubit.

  2. If using a USB drive, insert the USB drive into the instrument. If using a computer, connect the Qubit™ instrument to the computer using a USB cable. The Qubit™
    Fluorometer will be recognized as an external device by the computer.
    Note: The connection between the computer and the Qubit™ instrument  must be maintained until the transfer and update (described in the next steps) are complete.
    a. On your computer, find the Qubit™ software update file that you wish to upload.
    b. Copy and paste or click-drag the software file to the Qubit™ instrument drive. This saves the software file to the root directory of your Qubit™ instrument; however, the software file transfer will not be visible on the Qubit™ instrument.

  3. On the Home screen, press Settings.

  4. On the Settings screen, press Update software. The instrument searches the USB drive or the computer for the update.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
71 Note: If the USB drive is not inserted into the USB drive port or the instrument does not recognize the USB drive, a warning message is displayed.
    To proceed with the software update, insert the correct USB drive into the instrument, then press Close.

  5. When the appropriate files are detected, the Software update screen is displayed. Press Update to update the software.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  6. When prompted, press Restart to complete the software update.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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System verification test

Perform System verification test

The system verification test checks the internal components of the Qubit™ 4 Fluorometer and requires the use of the Qubit™ 4 System Verification Assay Kit (Cat. No. Q33237). Perform the system verification when a problem with the instrument is suspected. It is not necessary to regularly perform the verification.

Required materials not provided:

  • Qubit™ 4 System Verification Assay Kit (Cat. No. Q33237)
  • Qubit™ assay tubes (500 tubes, Cat. No. Q32856)
  1. On the Home screen, press Settings then System Verification Test. Press Start Test to proceed.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
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  2. Set up 3 Qubit™ assay tubes (Cat. No. Q32856).

  3. Label the tube lids 1, 2, and 3.
    Note: Do not label the side of the tube as this could interfere with the sample read.

  4. Add 200 µL of the Blank Reagent (Component A) to Tube 1.

  5. Add 200 µL of the Green Fluorescence Reagent (Component B) solution to Tube 2.

  6. Add 200 µL of the Far Red Fluorescence Reagent (Component C) solution to Tube 3.

  7. Insert Tube 1 containing the Blank Reagent (Component A) into the sample chamber, close the lid, then press Read tube. When the reading is complete (~3 seconds), remove Tube 1.

  8. Insert Tube 2 containing the Green Fluorescence Reagent (Component B) into the Qubit™ sample chamber, close the lid, then press Read tube. When the reading is complete (~3 seconds), remove Tube 2.

  9. Insert Tube 3 containing the Far Red Fluorescence Reagent (Component C) into the sample chamber, close the lid, then press Read tube. When the reading is complete (~3 seconds), remove Tube 3.
    Note: Always insert tubes in the order 1, 2, 3.

  10. Upon read completion, the instrument displays the overall result of the system verification.

  • If no errors are found, the System Verification Pass message appears.
    Press Close to return to the Settings screen or press Next to view the System Verification Report.
    Thermo Fisher Scientific MAN0017209 Qubit 4 Fluorometer With WiFi - Fig
75

  • If errors are found, the Error Reading Reagents message appears.
    Verify that the test was run with the lid closed, then press OK to rerun the test with the tube strips in the correct order.

  • If the System Verification Failed message persists after re-running the tube strips with the lid closed, contact Technical Support for help (page 70).

  • Press Next to view system verification test results details or press Data Log to access and export system verification test results.

Appendix A: Troubleshooting

Handling samples

  • The calibration standards included in the Qubit™ microRNA, Qubit™ RNA HS, Qubit™ RNA XR, Qubit ™ RNA IQ, and Qubit™ RNA BR Assay Kits are high- quality RNA standards. The integrity and concentration of these standards is critical to the optimal performance of the Qubit™ RNA assays. We highly recommend treating the rRNA standards as you would any other precious RNA. Use appropriate RNAse-free handling techniques, including RNAsefree gloves, pipette tips, and tubes. Keep the tube lids closed whenever possible; do not press the pipet  tothe inside wall of  the tube when withdrawing a sample. Return the RNA standards to –80°C as soon as possible after use.
  • Ensure that the assay tubes are at room temperature at the time the reading is taken. Do not hold assay tubes in your hand and do not leave assay tubes in the Qubit™ 4  Fluorometer for longer than it takes to read the fluorescence. See “Assay temperature”, page 60.
  • Be careful not to spill sample into the sample chamber. Promptly wipe any spills.
  • The Qubit™ assays are very sensitive and even small amounts of material from a previous sample may result in errors. Use a clean 0.5-mL PCR tube for each reading.
  • The tube must be clean and dry on the outside when taking readings. Moisture and condensation on the tube surface can lead to reading errors.
  • Minute bubbles in samples will cause errors in readings. Be sure not to introduce bubbles into samples. Slight tapping on the tube wall or brief centrifugation will often help dissipate bubbles.

High reading

  • The sample is out of range. Use a sample that is less concentrated.
  • For Qubit™ quantitation assays, view the Fluorescence vs. Concentration graph in the Results screen to confirm that the values for the samples fall between the values of  the standards (page 27).
  • Ensure that the lid is closed while reading standards and samples.
  • Prepare samples and standards according to the instructions in the Qubit™ assay kit you are using.
  • Ensure that the assay is performed entirely at room temperature.

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Low reading

  • The sample is out of range. Use a sample that is more concentrated or use a lower dilution (for example, 20 µL in 180 µL instead of 10 µL in 190 µL).
  • For Qubit™ quantitation assays, view the Fluorescence vs. Concentration graph in the Results screen to confirm that the values for the samples fall between the values of the standards (page 27).
  • Ensure that you have prepared the Qubit™ working solution correctly (1:200 dilution using the buffer provided in the kit).
  • Ensure that you have prepared the standard tubes correctly (10 µL of each standard in 190 µL of Qubit™ working solution).
  • Ensure that the standard and sample tubes are filled to 200 µL.
  • Protect the Qubit™ reagent and working solutions from light.
  • Select the correct Qubit™ 4 Fluorometer assay for the Qubit™ assay you are performing and calibrate the fluorometer correctly. Standards must be used in the correct order.
  • Ensure that the assay is performed entirely at room temperature.

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Critical Qubit™ Assay considerations

How the Qubit™ 4 Fluorometer calculates concentration

The Qubit™ 4 Fluorometer generates concentration data based on the relationship between the two standards used in calibration (three for the Qubit™ protein assay). The plot below shows the line corresponding to the curve-fitting algorithm (a modified Hill plot) used in the calculation of concentration data for the Qubit™ RNA HS assay. For reference, the positions of the standards and a set of data points from an actual experiment are shown superimposed onto the line. This plot demonstrates that the curve-fitting algorithm gives accurate values for quantitation.

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Figure 1. The curve-fitting algorithm used to determine concentration in the Qubit™ RNA HS assay. Data for other Qubit™ quantitation assays are generated by similar algorithms.

Incubation time

To allow the Qubit ™ assay to reach maximum fluorescence, incubate the tubes for the DNA and RNA assays for 2 minutes after mixing the sample or standard with the working solution. After this incubation period, the fluorescence signal is stable for 3 hours at room temperature for all nucleic acid assays except the Qubit ™ ssDNA assay, which is stable for up to 30 minutes. The Qubit ™ protein assay requires 15 minutes of incubation for a stable signal. For greatest accuracy in the protein assay, the incubation time of the samples should be within 10 minutes of the incubation time of the standards. The Qubit ™ Protein Broad Range Assay requires 10 minutes of incubation time for best results.

Photobleaching of Qubit™ reagents

The Qubit ™ DNA and protein exhibit high photostability in the Qubit ™ 4 Fluorometer, showing <0.3% drop in fluorescence after 9 readings and <2.5% drop in fluorescence after 40 readings. It is important to remember, however, that if the assay tube remains in the Qubit ™ 4 Fluorometer for multiple readings, a temporary reduction in fluorescence will be observed as the solution increases in temperature (see Figure 2 in “Assay temperature”, page 60). The RNA assays including the RNA IQ assay should only be read once. Note that the temperature inside the Qubit ™ 4 Fluorometer may be as much as 3°C above room temperature after 1 hour. For this reason, if you want to perform multiple readings of a single tube, remove the tube from the instrument and let it equilibrate to room temperature for 30 seconds before taking another reading.

Assay temperature

The Qubit ™ assays were designed to be performed at room temperature (22–28°C), and temperature fluctuations can influence the accuracy of the assay. To minimize temperature fluctuations, store all kit reagents at room temperature and insert all assay tubes into the Qubit ™ 4 Fluorometer only for as much time as it takes for the instrument to measure the fluorescence, because the Qubit ™ 4 Fluorometer can raise the temperature of the assay solution significantly, even over a period of a few minutes. Do not hold the assay tubes in your hand before a measurement, because holding the tubes warms the solution and results in a low reading.

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Figure 2. Effect of temperature on the Qubit™ dsDNA BR assay. Qubit™ dsDNA HS, Qubit™  ssDNA, Qubit ™ RNA HS, Qubit™ RNA XR, Qubit  ™ RNA IQ, and  Qubit ™ protein assays show similar sensitivities over the same range.

Qubit™ 4 Fluorometer calibration

For each assay, you have the choice to run a new calibration or to use the values from the previous calibration. As you first use the instrument, you perform a new calibration each time. As you become familiar with the assays, the instrument, your pipetting accuracy, and significant temperature fluctuations within your laboratory, you can determine the level of comfort you have using the calibration data stored from the last time the instrument was calibrated. Remember also that the fluorescence signal in the tubes containing the standards and the samples is stable for not longer than 3 hours. See Figure 1 in “How the Qubit ™ 4 Fluorometer calculates concentration” (page 59) for an example of the calibration curve used to generate the quantitation results.

Appendix B: Ordering information

Qubit™ 4 Fluorometer and accessories
The following products can be used with the Qubit™ 4 Fluorometer and are available separately from Thermo Fisher Scientific. For more information, visit thermofisher.com or contact Technical Support (page 71).

Product Quantity Cat. No.
QubiC 4 Fluorometer, with Wi-Fi 1 kit Q33238
Qubitna 4 Quantitation Starter Kit, with Wi-Fi 1 kit Q33239
Qubitne 4 NGS Starter Kit, with Wi-Fi 1 kit Q33240
Qubitim RNA IQ Starter Kit, with Wi-Fi 1 kit Q33241
QubiC 4 USB Flash Drive 1 each Q46009
QubiC 4 Fluorometer International Power Supply (replacement) 1 each A36204
Qubitim Assay Tubes 500 tubes Q32856
100 assays A50668
Qubitim Protein BR Assay Kit 111-20 mg/mL 500 assays A50669
100 assays Q33211
Qubit”‘ Protein Assay Kit 0.25-5 pg 500 assays Q33212
Qubir 1X dsDNA HS Assay Kit 100 assays
500 assays Q33230

Q33231
Qubir lx dsDNA BR Assay Kit| 100 assays
500 assays| Q33265
Q33266
Qubitna ssDNA Assay Kit 1-200 ng| 100 assays| Q10212
Qubir RNA IQ Assay Kit| 75 assays
275 assays| Q33221
Q33222
Qubir RNA HS Assay Kit| 100 assays
500 assays| Q32852
Q32855
Qubir RNA BR Assay Kit| 100 assays
500 assays| Q10210
Q10211
Qubir RNA XR Assay Kit| 100 assays
500 assays| Q33223
Q33224
QubiC microRNA Assay Kit 0.5-100 ng| 100 assays
500 assays| Q32880
Q32881

Appendix C: Safety

Symbols on instruments
Electrical symbols The following table describes the electrical symbols that may be displayed.

Symbol Description
**** Indicates a terminal that can receive or supply alternating current or

voltage.

Safety symbols

The following table describes the safety symbols that may be displayed. Each symbol may appear by itself or in combination with text that explains the relevant hazard (see “Safety labels on instruments”). These safety symbols may also appear next to DANGERS, WARNINGS, and CAUTIONS that occur in the text of this and other product-support documents.

Symbol Description
Indicates that you should consult the manual for further information and to

proceed with appropriate caution.
| Indicates the presence of an electrical shock hazard and to proceed with
appropriate caution.

Environmental symbols

The following symbol applies to all Thermo Fisher Scientific electrical and electronic products placed on the European market after August 13, 2005.

Symbol Description
Do not dispose of this product as unsorted municipal waste. Follow local

municipal waste ordinances for proper disposal provisions to reduce the environmental impact of waste electrical and electronic equipment (WEEE).
European Union customers:
Call your Customer Service representative for equipment pick-up and recycling. See thermofisher.com for a list of customer service offices in the European Union.

Safety labels on instruments
The following CAUTION, WARNING, and DANGER statements may be displayed on Thermo Fisher Scientific instruments in combination with the safety symbols described in the preceding section.

symbol|
---|---
| CAUTION! Hazardous chemicals. Read the Safety Data Sheets (SDSs) before handling.
CAUTION! Hazardous waste. Refer to SDS(s) and local regulations for handling and disposal.
| DANGER! High voltage.
WARNING! To reduce the chance of electrical shock, do not remove covers that require tool access. No user-serviceable parts are inside. Refer servicing to Thermo Fisher Scientific qualified service personnel.

General instrument safety

WARNING! PHYSICAL INJURY HAZARD. Use this product only as specified in this document. Using this instrument in a manner not specified may result in personal injury or damage to the instrument.

Operating the instrument

Ensure that anyone who operates the instrument has:

  • Received instructions in both general safety practices for laboratories and specific safety practices for the instrument.
  • Read and understood all applicable Safety Data Sheets (SDSs). See “Safety Data Sheets (SDS)”.

Safety precautions

Do not install the instrument in heavy humidity such as a greenhouse or an incubator to avoid a danger of electric shock. If water or other material enters the instrument, the adaptor, or power inlet, disconnect the power cord and contact a service person. For operating environment, refer to “Product specifications” (page 8).

  • Do not press the main plug or power cord with wet hands.
  • Always ensure that the power supply input voltage matches the voltage available in your location.
  • Do not install the instrument on a slant or a place prone to vibrations, which induces the risk of instrument malfunction or damage of the instrument.
  • Plug the power cord firmly into the wall outlet and the instrument.
  • To avoid potential shock hazard, make sure that the power cord is properly grounded.
  • Be sure to position the equipment such that it is easy to disconnect the instrument.
  • If the instrument is broken or dropped, disconnect the power cord and contact technical services. Do not disassemble the instrument.
  • Use only authorized accessories (adaptor, power cord, and USB drive).
  • For operating environment, see “Product specifications” (page 8).
  • If the instrument emits smoke, disconnect the power cord from the wall outlet and contact technical services.

Cleaning or decontaminating the instrument
CAUTION! Using cleaning or decontamination methods other than those recommended by the manufacturer may compromise the safety or quality of the instrument.

Removing covers or parts of the instrument
CAUTION! PHYSICAL INJURY HAZARD The instrument is to be serviced only by trained personnel or vendor specified in the user guide. Do not remove any covers or parts that require the use of a tool to obtain access to moving parts.

Chemical safety
Chemical hazard warning

WARNING! CHEMICAL HAZARD. Before handling any chemicals, refer to the Safety Data Sheet (SDS) provided by the manufacturer, and observe all relevant precautions.
WARNING! CHEMICAL STORAGE HAZARD. Never collect or store waste in a glass container because of the risk of breaking or shattering. Reagent and waste bottles can crack and leak. Each waste bottle should be secured in a low- density polyethylene safety container with the cover fastened and the handles locked in the upright position. Wear appropriate eyewear, clothing, and gloves when handling reagent and waste bottles.

General safety guidelines
To minimize the hazards of chemicals:

  • Read and understand the Safety Data Sheets (SDSs) provided by the chemical manufacturer before you store, handle, or work with any chemicals or hazardous materials. (See “Safety Data Sheets (SDS)”, page 71)
  • Minimize contact with chemicals. Wear appropriate personal protective equipment when handling chemicals (for example, safety glasses, gloves, or protective clothing). For additional safety guidelines, consult the SDS.
  • Minimize the inhalation of chemicals. Do not leave chemical containers open. Use only with adequate ventilation (for example, fume hood). For additional safety guidelines, consult the SDS.
  • Check regularly for chemical leaks or spills. If a leak or spill occurs, follow the manufacturer’s cleanup procedures as recommended in the SDS.
  • Comply with all local, state/provincial, or national laws and regulations related to chemical storage, handling, and disposal.

Chemical waste safety
Chemical waste hazard
CAUTION! HAZARDOUS WASTE. Refer to Safety Data Sheets (SDSs) and local regulations for handling and disposal.
Chemical waste safety guidelines

To minimize the hazards of chemical waste:

  • Read and understand the Safety Data Sheets (SDSs) provided by the manufacturers of the chemicals in the waste container before you store, handle, or dispose of chemical waste. (See “Safety Data Sheets (SDS)”, page 71)
  • Provide primary and secondary waste containers. (A primary waste container holds the immediate waste. A secondary container contains spills or leaks from the primary container. Both containers must be compatible with the waste material and meet federal, state, and local requirements for container storage.)
  • Minimize contact with chemicals. Wear appropriate personal protective equipment when handling chemicals (for example, safety glasses, gloves, or protective clothing). For additional safety guidelines, consult the SDS.
  • Minimize the inhalation of chemicals. Do not leave chemical containers open. Use only with adequate ventilation (for example, fume hood). For additional safety guidelines, consult the SDS.
  • Handle chemical wastes in a fume hood.
  • After emptying the waste container, seal it with the cap provided.
  • Dispose of the contents of the waste tray and waste bottle in accordance with good laboratory practices and local, state/provincial, or national environmental and health regulations.

Waste disposal

If potentially hazardous waste is generated when you operate the instrument, you must:

  • Characterize (by analysis, if necessary) the waste generated by the particular applications, reagents, and substrates used in your laboratory.
  • Ensure the health and safety of all personnel in your laboratory.
  • Ensure that the instrument waste is stored, transferred, transported, and disposed of according to all local, state/provincial, and/or national regulations.

IMPORTANT! Radioactive or biohazardous materials may require special handling, and disposal limitations may apply.

Electrical safety

DANGER! ELECTRICAL SHOCK HAZARD. Severe electrical shock can result from operating the Qubit™ 4 Fluorometer without its instrument panels in place. Do not remove instrument panels. High‐voltage contacts are exposed when instrument panels are removed from the instrument.
Power
DANGER! ELECTRICAL HAZARD. Grounding circuit continuity is vital for the safe operation of equipment. Never operate equipment with the grounding conductor disconnected.
DANGER! ELECTRICAL HAZARD. Use properly configured and approved line cords for the voltage supply in your facility.
DANGER! ELECTRICAL HAZARD. Plug the system into a properly grounded receptacle with adequate current capacity.

Overvoltage rating
The Qubit™ 4 Fluorometer has an installation (overvoltage) category of II, and is classified as portable equipment.

Biological hazard safety
WARNING!
BIOHAZARD. Biological samples such as tissues, body fluids, and blood of humans and other animals have the potential to transmit infectious diseases. Follow all applicable local, state/provincial, and/or national regulations. Wear appropriate protective eyewear, clothing, and gloves. Read and follow the guidelines in these publications.

In the U.S.:

In the EU:

Safety, Electromagnetic compatibility (EMC), and Environmental standards

This section provides information on:

  • U.S. and Canadian safety and EMC standards
  • European safety, EMC, and Environmental standards
  • Australian EMC standards
  • China RoHS Standards

U.S. and Canadian Safety standards

The Qubit™ 4 Fluorometer has been tested to and complies with standard: UL 61010-1/CAN/CSA-C22.2 No. 61010-1, “Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use, Part 1: General Requirements.”
UL 61010-2-081/CAN/CSA-C22.2 No. 61010-2-081, “Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use, Part 2-081: Particular Requirements for Automatic and Semi-Automatic Laboratory Equipment for Analysis and Other Purposes.”

U.S. EMC standard

This instrument has been tested to and complies with standard 47 CFR FCC Part 15 “Radio Frequency Devices”; Subpart B “Unintentional Radiators”.
Note: This equipment has been tested and found to comply with the limits for a Class B digital device, pursuant to part 15 of the FCC Rules. These limits are designed to provide reasonable protection against harmful interference in a residential installation. This equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in accordance with the instructions, may cause harmful interference to radio communications. However, there is no guarantee that interference will not occur in a particular installation. If this equipment does cause harmful interference to radio or television reception, which can be determined by turning the equipment off and on, the user is encouraged to try to correct the interference by one or more of the following measures:

  • Reorient or relocate the receiving antenna.
  • Increase the separation between the equipment and receiver.
  • Connect the equipment into an outlet on a circuit different from that to which the receiver is connected.
  • Consult the dealer or an experienced radio/TV technician for help.

Canadian EMC standard

This instrument has been tested to and complies with standard ICES-001, “Industrial, Scientific and Medical (ISM) Radio Frequency Generators.”

European Safety, EMC, and Environmental standards

Safety
This instrument meets European requirements for safety (Low Voltage Directive 2014/35/EU). This instrument has been tested to and complies with standards:
IEC/EN 61010-1:2010, “Safety Requirements for Electrical Equipment for Measurement, Control and Laboratory Use, Part 1: General Requirements.”
IEC/EN 61010-2-081:2015, “Safety requirements for electrical equipment for measurement, control, and laboratory use – Part 2-081: Particular requirements for automatic and semi-automatic laboratory equipment for analysis and other purposes.”

EMC
This instrument meets European requirements for emission and immunity (EMC Directive 2014/30/EU). This instrument has been tested to and complies with standards:
EN 61326-1:2013 Class B, “Electrical Equipment for Measurement, Control and Laboratory Use – EMC Requirements.”
EN 61000-3-2:2014, “Electromagnetic compatibility (EMC) — Part 3 – 2: Limits — Limits for harmonic current emissions (equipment input current ≤16 A per phase).”
EN 61000-3-3:2013, “Electromagnetic compatibitity (EMC) — Part 3 – 3: Limits – Limitation of voltage changes, voltage fluctuations and flicker in public lowvoltage supply
systems, for equipment with rated current ≤ 16 A per phase and not subject to conditional connection (IEC 61000‒3‒3:2008).”

Restriction of Hazardous Substance (RoHS)
This instrument meets European requirements RoHS Directive 2011/65/EU.
This instrument has been tested to and complies with standard EN 50581:2012, “Technical documentation for the evaluation of electrical and electronic products with respect to restriction of hazardous substances.”

Waste of Electrical and Electronic Equipment (WEEE)
This instrument meets European requirement WEEE Directive 2012/19/EU.

Australian EMC standards

This instrument has been tested to and complies with standard AS/NZS CISPR 11, “Limits and Methods Measurement of Electromagnetic Disturbance Characteristics of Industrial, Scientific, and Medical (ISM) Radio‐frequency Equipment.”

China RoHS standards

This instrument does not contain any hazardous substances above the limit and complies with China RoHS standards:
GB/T 26572-2011, “Requirements on Concentration Limits for Certain Restricted Substances in Electrical and Electronic Products”
SJ/T 11364-2014, “Marking for the Restricted Use of Hazardous Substances in Electronic and Electrical Products”

Documentation and support

Obtaining support
Technical support

Visit thermofisher.com/support for the latest in services and support, including:

  • Worldwide contact telephone numbers

  • Product support, including:
    – Product FAQs
    – Software, patches, and updates
    – Training for many applications and instruments

  • Order and web support

  • Product documentation, including:
    – User guides, manuals, and protocols
    – Certificates of Analysis
    – Safety Data Sheets (SDSs; also known as MSDSs)

Safety Data Sheets (SDS)
Safety Data Sheets (SDSs) are available at thermofisher.com/support.

IMPORTANT! For SDSs for reagents and chemicals from other manufacturers, contact the manufacturer.

Limited product warranty

Life Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies’ General Terms and Conditions of Sale found on Life Technologies’ website at www.thermofisher.com/us/en/home/global /termsand-conditions.html. If you have any questions, please contact Life Technologies at www.thermofisher.com/support.
IMPORTANT! Wiping the Qubit™ 4 Fluorometer computer (i.e., erasing  the hard drive to remove all programs, files, and the operating system) voids the product warranty.

thermofisher.com/support | thermofisher.com/askaquestion
thermofisher.com
20 October 2021

References

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