Promega AS1400 Maxwell RSC Blood DNA Kit User Manual
- June 3, 2024
- Promega
Table of Contents
TECHNICAL MANUAL
Maxwell® RSC Blood DNA Kit
Instructions for Use of Products
AS1400 and ASB1400
AS1400 Maxwell RSC Blood DNA Kit
Caution: Handle cartridges with care; seal edges may be sharp.
All technical literature is available at:
www.promega.com/protocols/
Visit the web site to verify that you are using the most current version of
this Technical Manual.
E-mail Promega Technical Services if you have questions on use of this system:
techserv@promega.com
Description
The Maxwell ® RSC Blood DNA Kit (a) is used with the Maxwell ® RSC or Maxwell
® RSC 48 Instruments to provide an easy method for efficient, automated
purification of genomic DNA (gDNA) from samples. The Maxwell ® RSC and Maxwell
® RSC 48 Instruments are supplied with preprogrammed purification procedures
and are designed for use with predispensed reagent cartridges, maximizing
simplicity and convenience. The Maxwell ® RSC Instrument can process up to 16
samples and the Maxwell ® RSC 48 instrument can process up to 48 samples. Both
instruments can process the maximum number of samples in about 40 minutes. The
purified DNA can be used directly in a variety of downstream applications.
The Maxwell ® RSC Blood DNA Kit purifies samples using a novel paramagnetic
particle, called the MagnaCel™ particle, which provides a mobile solid phase
that optimizes sample capture, washing and purification of gDNA. This particle
utilizes cellulose-based binding of nucleic acids and provides a higher bind
capacity and cleaner eluate than traditional DNA purification. The Maxwell ®
RSC and Maxwell ® RSC 48 are magnetic particle-handling instruments that
efficiently bind gDNA to the paramagnetic particle in the first well of a
prefilled cartridge. The samples are processed through a series of washes
before the gDNA is eluted.
Description (continued)
Prior to extraction, samples can be preprocessed manually or using the
Maxprep™ Liquid Handler. The Maxprep™ Liquid Handler will scan bar codes and
transfer samples from primary blood tubes, perform sample lysis prior to
extraction, transfer lysed samples to Maxwell ® RSC Blood DNA cartridges, add
plungers to Maxwell® RSC Blood DNA cartridges, and add Elution Buffer to
elution tubes. Instructions are provided for preprocessing samples manually or
with the Maxprep™ Liquid Handler.
Product Components and Storage Conditions
PRODUCT | SIZE | CAT.# |
---|---|---|
Maxwell® RSC Blood DNA Kit | 48 preps | AS1400 |
For Research Use. Sufficient for 48 automated isolations from up to 300µl of whole blood samples. Includes:
- 2 × 1ml Proteinase K (PK) Solution
- 20ml Lysis Buffer
- 48 Maxwell® RSC Cartridges
- 1 Maxwell ® RSC Plunger Pack (48 Plungers)
- 50 Elution Tubes (0.5ml)
- 20ml Elution Buffer
PRODUCT | SIZE | CAT.# |
---|---|---|
Maxwell® RSC Blood DNA Multi-Pack Kit | 144 preps | ASB1400 |
For Research Use. Sufficient for 144 automated isolations from up to 300µl of
whole blood samples.
Note: ASB1400 is not recommended for use with the Maxprep™ Liquid
Handler. Includes:
- 6 × 1ml Proteinase K (PK) Solution
- 3 × 20ml Lysis Buffer
- 144 Maxwell ® RSC Cartridges
- 3 × 50/pk Maxwell ® CSC/RSC Plungers
- 3 × 50 Elution Tubes (0.5ml)
- 3 × 20ml Elution Buffer
Storage Conditions: Store the Maxwell ® RSC Blood DNA Kit at +15°C to +30°C.
Safety Information: The Maxwell ® RSC Cartridges contain ethanol and
isopropanol. These substances should be considered flammable, harmful and
irritants. Lysis Buffer contains guanidine hydrochloride and urea. These
substances should be considered toxic, harmful and irritants.
Maxwell ® RSC Cartridges are designed to be used with potentially infectious
substances. Wear appropriate protection (e.g., gloves and goggles) when
handling potentially infectious substances. Adhere to your institutional
guidelines for the handling and disposal of all potentially infectious
substances used with this system. 6422MA
Available Separately (recommended for sample extraction)
For Manual Preprocessing
PRODUCT | SIZE | CAT.# |
---|---|---|
ClickFit Microtube, 1.5ml | 1,000/pack | V4741 |
For Preprocessing with the Maxprep™ Liquid Handler
PRODUCT | SIZE | CAT.# |
---|---|---|
2.0ml Deep Well Plates (Non-Sterile) | 60/pack | AS9309 |
Nunc 2.0ml Deep Well Plates (Sterile) | 60/pack | AS9307 |
Maxprep™ 1000µl Conductive Disposable Tips, Filtered | 40/box | AS9303 |
Maxprep™ 300µl Conductive Disposable Tips, Filtered | 60/box | AS9302 |
Maxprep™ Reagent Reservoir, 50ml | 28/pack | AS9304 |
Maxprep™ Plunger Holder | 1 each | AS9408 |
Maxwell® RSC Plunger Pack | 48/each | AS1670 |
Maxprep™ 3-Position Reagent Tube Holder | 1 each | AS9409 |
Manual Peparation of Blood Samples
Materials to Be Supplied by the User
- optional, rotating tube mixer for liquid blood samples
- benchtop vortex mixer
- pipettors and pipette tips for sample transfer into prefilled reagent cartridges
- 1.5–2.0ml tubes for incubation of samples (e.g., ClickFit Microtube, 1.5ml [Cat.# V4741]; recommended to prevent the cap from opening during heating)
- heating block set at 56°C
The total yield of genomic DNA from whole blood samples depends on the sample
volume and number of white blood cells/ml. Each Maxwell ® RSC Cartridge
supplied in the Maxwell ® RSC Blood DNA Kit is designed to purify genomic DNA
from up to 300µl of whole blood, assuming an average number of white blood
cells in the range of 4 × 10 6 to 1.1 × 10 7 cells/ml whole blood (values for
a normal healthy adult; 1).
Note: Whole blood samples collected in EDTA, ACD or heparin tubes can be
used. These samples may be either fresh or frozen. Frozen samples must be
thawed before processing.
3.A. Preparation of Whole Blood Samples
-
Mix all blood samples for at least 5 minutes at room temperature.
-
Prepare and label incubation tubes compatible with a heating block.
-
Add 30µl of Proteinase K (PK) Solution to each incubation tube.
-
Add liquid blood (up to 300µl) to each incubation tube.
3.A. Preparation of Whole Blood Samples (continued) -
Add 300µl of Lysis Buffer to each incubation tube.
-
Vortex each tube for 10 seconds.
-
Incubate each tube in the heating block (set to 56°C) for 20 minutes. During this incubation, prepare cartridges as described in Section 3.B.
-
Transfer each blood lysate sample from the incubation tube to well #1 of each cartridge. (Well #1 is the well closest to the printed side and furthest from the elution tube.)
3.B. Maxwell ® RSC Blood DNA Cartridge Preparation
- Change gloves before handling Maxwell® RSC Cartridges, RSC Plungers and Elution Tubes (0.5ml). Place each cartridge in the deck tray with the printed side facing away from the elution tubes. Press down on both the front and back of the cartridge to snap it into position. Carefully peel back the seal so that all plastic comes off the top of the cartridge. Ensure that all sealing tape and any residual adhesive are removed before placing cartridges in the instrument.
- Place one plunger into well #8 of each cartridge.
- Place an empty elution tube into the elution tube position for each cartridge in the deck tray. Add 50µl of Elution Buffer to the bottom of each elution tube.
- Proceed to Section 5 for Maxwell ® Instrument setup and run.
Notes:
- Specimen or reagent spills on any part of the deck tray should be cleaned with a detergent-water solution, followed by a bacteriocidal spray or wipe, then water. Do not use bleach on any instrument parts.
- Use only the 0.5ml Elution Tubes provided in the kit; other tubes may be incompatible with the Maxwell® Instrument.
User Adds to Wells
1. Sample lysate
8. RSC Plunger
Figure 1. Maxwell® RSC Cartridge.
Figure 2. Setup and configuration of the deck tray. Elution Buffer is added to
the elution tubes as shown. Plungers are in well #8 of the cartridge.
Maxprep™ Preprocessing
Materials to Be Supplied by the User
- optional, rotating tube mixer for liquid blood samples
The total yield of genomic DNA from whole blood samples depends on the sample volume and number of white blood cells/ml. Each Maxwell ® RSC Cartridge supplied in the Maxwell ® RSC Blood DNA Kit is designed to purify genomic DNA from up to 300µl of whole blood, assuming an average number of white blood cells in the range of 4 × 10 6 to 1.1 × 10 7 cells/ml whole blood (values for a normal healthy adult; 1). Note: Whole blood samples collected in EDTA, ACD or heparin tubes can be used. These samples may be either fresh or frozen. Frozen samples must be thawed before processing.
4.A. Maxprep™ Cartridge Preparation
-
Mix all blood samples for at least 5 minutes at room temperature.
-
Turn on the Maxprep™ Liquid Handler and PC. Log in to the PC, and start the Maxprep™ software on the PC by double-clicking the desktop icon.
-
Press Start to access the ‘Methods’ screen.
-
On the ‘Methods’ screen, select a method using one of the two options below:
1. Touch the Blood DNA preprocessing method or laboratory-specific variant of the Blood DNA preprocessing method.
2. Use a bar code reader to scan the 2D bar code on the kit box to automatically select the appropriate base method. Touch the laboratory- specific user variant of the Blood DNA preprocessing method if desired. -
Verify that the appropriate preprocessing method or user-variant method has been selected, and press the Proceed button. Touch the Run button on the method run screen to start the run.
-
Enter any method-specific variables (Sample Number, Sample Volume, Elution Volume).
4.A. Maxprep™ Cartridge Preparation (continued) -
Prior to placing Maxwell ® RSC or Maxwell ® RSC 48 deck trays on the Maxprep™ Liquid Handler, prepare the deck trays with cartridges and elution tubes. Change gloves before handling Maxwell ® RSC Cartridges, RSC Plungers and Elution Tubes (0.5ml). Place each cartridge in the deck tray(s) with the printed side facing away from the elution tubes. Press down on both the front and back of the cartridge to snap it into position. Carefully peel back the seal so that all plastic comes off the top of the cartridge. Ensure that all sealing tape and any residual adhesive are removed before placing cartridges in the Maxprep™ Liquid Handler. Place an empty elution tube into the elution tube position for each cartridge in the deck tray(s).
Notes:
1. Specimen or reagent spills on any part of the deck tray should be cleaned with a detergent-water solution, followed by a bacteriocidal spray or wipe, then water. Do not use bleach on any instrument parts.
2. Use only the 0.5ml Elution Tubes provided in the kit; other tubes may be incompatible with the Maxwell ® Instruments. -
Follow instrument setup instructions displayed in the method. The Maxprep™ software will direct you where to place the following items on the instrument:
Labware Type
• Maxprep™ Plunger Holders and Maxwell ® RSC Plunger Packs (2; one may be partially full)
• Maxwell ® RSC 48 Front deck tray or Maxwell ® RSC deck tray containing Maxwell ® RSC cartridges with seals removed and open elution tubes
• Maxwell ® RSC 48 Back deck tray or Maxwell ® RSC deck tray containing Maxwell ® RSC cartridges with seals removed and open elution tubes (if you are running greater than 24 samples in a run)
• Maxprep™ 3-Position Reagent Tube Holder with up to 3 Proteinase K tubes
• Maxprep™ Reagent Reservoir, 50ml, with Lysis Buffer
• Maxprep™ Reagent Reservoir, 50ml, with Elution Buffer
• Nunc 2.0ml Deep-well Plate (empty)
• Maxprep™ 1000μl Conductive Disposable Tips, Filtered (2; one rack may be partially full)
• Maxprep™ 300μl Conductive Disposable Tips, Filtered (rack may be partially full) -
Close the Maxprep™ Liquid Handler door, and touch the Next button to start the automated preprocessing of your samples.
4.B. Maxprep™ Liquid Handler Preprocessing Protocol
The Maxprep™ Liquid Handler will use the following steps to prepare samples
prior to extraction with the Maxwell® RSC or Maxwell ® RSC 48 Instrument.
-
The system prepares a lysis reaction in the Nunc 2.0ml Processing Plate consisting of the following components:
a. The specified volume of whole blood
b. 30µl of Proteinase K Solution
c. 300µl of Lysis Buffer -
The Processing Plate incubates for 20 minutes.
-
During the lysis incubation, plungers are transferred to each of the cartridges in the Maxwell ® RSC or Maxwell ® RSC 48 deck trays. The specified volume of Elution Buffer is transferred to the elution tubes for each position in the Maxwell ® RSC or Maxwell ® RSC 48 deck tray.
-
After lysis incubation is complete, each sample is transferred from the Processing Plate to its corresponding Maxwell ® RSC cartridge.
-
Once the method is complete, open the Maxprep™ Liqiud Handler door, and transfer the deck trays to your Maxwell ® RSC or Maxwell ® RSC 48 Instrument for extraction. Remove primary sample tubes and Processing Plate, and discard the used tips from the waste bin as hazardous waste following your institution’s recommended guidelines. Either discard or tightly cap and store remaining reagents.
Consumables used by the Maxprep™ Instrument for preprocessing methods are designed to be used with potentially infectious substances. Use appropriate protective equipment (e.g., gloves and goggles) when handling infectious substances. Adhere to your institutional guidelines for the handling and disposal of all infectious substances when used with this system.
Maxwell ® RSC Instrument Setup and Run
Refer to the Maxwell ® RSC Instrument Operating Manual #TM411 or the Maxwell ® RSC 48 Instrument Operating Manual #TM510 for detailed information.
-
Turn on the Maxwell ® Instrument and Tablet PC. Log in to the Tablet PC, and start the maxwell ® Software by double-touching the icon on the desktop. The instrument will power up, proceed through a self-check and home all moving parts.
-
Touch Start to access the ‘Methods’ screen.
When running in Portal mode, scan the bar code(s) on the deck tray(s). After data has been returned from the Portal database, press Continue to use the sample tracking information for the deck tray(s) or press New to start a run and enter new sample tracking information. -
On the ‘Methods’ screen, if a method has not been chosen by scanning the bar code on the deck trays, select a method using one of the two options below: a. Touch the RSC Blood DNA method. b. Use a bar code reader to scan the 2D bar code on the kit box to automatically select the appropriate method.
-
Verify that the RSC Blood DNA method has been selected, and press the Proceed button. If requested by the software, enter any kit lot and expiration information that has been required by the Administrator.
-
On the ‘Cartridge Setup’ screen (if shown), touch the cartridge positions to sselect/deselect any positions to be used for this extraction run. Enter any required sample tracking information, and press the Proceed button to continue.
Note: When using the Maxwell ® RSC 48 Instrument, use the Front and Back buttons to select/deselect cartridge positions on each deck tray. -
After the door has been opened, confirm that all Extraction Checklist items have been performed. Verify that samples were added to well #1 of the cartridges, cartridges are loaded on the instrument, uncapped elution tubes are present with Elution Buffer and plungers are in well #8. Transfer the deck tray(s) containing the prepared cartridges onto the Maxwell ® instrument platform. Inserting the Maxwell ® deck tray(s): Hold the deck tray by the sides to avoid dislodging cartridges from the deck tray. Ensure that the deck tray is placed in the Maxwell ® Instrument with the elution tubes closest to the door. Angle the back of the deck tray downward and place into the instrument so that the back of the deck tray is against the back of the instrument platform. Press down on the front of the deck tray to firmly seat the deck tray on the instrument platform. If you have difficulty fitting the deck tray on the platform, check that the deck tray is in the correct orientation. Ensure the deck tray is level on the instrument platform and fully seated. Note: When using the Maxwell ® RSC 48 Instrument, check the identifier on the Maxwell ® RSC 48 deck tray to determine whether it should be placed in the front or back of the instrument.
-
Touch the Start button to begin the extraction run. The platform will retract, and the door will close.
Warning: Pinch point hazard.
The Maxwell ® Instrument will immediately begin the purification run. The screen will display information including the user who started the run, the current method step being performed, and the approximate time remaining in the run.
Notes:
1. When using the Maxwell ® RSC 48 Instrument, if the Vision System has been enabled, the deck trays will be scanned as the door retracts. Any errors in deck tray setup (e.g., plungers not in well #8, elution tubes not present and open) will cause the software to return to the ‘Cartridge Setup’ screen and problem positions will be marked with an exclamation point in a red circle. Resolve all error states, and press the Start button again to repeat deck tray scanning and begin the extraction run.
2. Pressing the Abort button will abandon the run. All samples from an aborted run will be lost.
3. If the run is abandoned before completion, you may be prompted to check whether plungers are still loaded on the plunger bar. If plungers are present on the plunger bar, you should perform Clean Up when requested. If plungers are not present on the plunger bar, you can choose to skip Clean Up when requested. The samples will be lost. -
Follow on-screen instructions at the end of the method to open the door. Verify that plungers are located in well #8 of the cartridge at the end of the run. If plungers are not removed from the plunger bar, follow the instructions in the Maxwell ® RSC Instrument Operating Manual or the Maxwell ® RSC 48 Instrument Operating Manual to perform a Clean Up process to attempt to unload the plungers.
-
Remove the deck tray(s) from the instrument. Remove elution tubes containing DNA, and cap the tubes. After the run has been completed, the extraction run report will be displayed. From the “Report View’ screen, you can print or export this report or both.
-
Remove the cartridges and plungers from the deck tray, and discard as hazardous waste following your institution’s recommended guidelines. Do not reuse reagent cartridges, plungers or elution tubes.
Ensure samples are removed before performing any required UV light treatment to avoid damage to the nucleic acid. 6422MA
Reference
- Henry, J.B. (2001) Clinical Diagnosis and Management by Laboratory Methods, 20th ed., W.B. Saunders Company, 509.
Troubleshooting
For questions not addressed here, please contact your local Promega Branch Office or Distributor. Contact information available at: www.promega.com. E-mail: techserv@promega.com
Symptoms | –Causes and Comments |
---|---|
Lower than expected A2. (lower than expected yield) | Blood that has undergone |
multiple freeze-thaw cycles may have degraded DNA. Use fresh samples whenever
possible, or avoid multiple freeze-thaw cycles.
Proteinase K Solution was not added. The lysis and yield are dependent upon
complete extraction with Proteinase K. If Proteinase K was not added in
Section 3.A, Step 3, or set up on the MaxprepTh Liquid Handler, the resulting
blood sample will be red. Proteinase K-treated samples turn greenish brown.
This can be used as a quick diagnostic method of determining whether or not
Proteinase K was added.
Whole blood sample contained low white blood cell count. The yield of genomic
DNA from blood samples depends on the number of white blood cells present in
the sample
Whole blood sample was not mixed before processing. Be sure to mix whole blood
samples before processing to ensure that the white blood cells are in
suspension.
The Maxwell® RSC or RSC 48 Instrument was set for the wrong method. Ensure
that the Blood DNA method is chosen.
RNA contamination in DNA eluates| In some cases, total RNA can be copurified
with the genomic DNA. To remove copurified RNA, an RNase treatment can be
performed. Add 5111 of RNase A (Cat.* A7973) per milliliter of Elution Buffer.
Instrument unable to pick up plungers| Make sure you are using an RSC-specific
chemistry kit; the plungers for the Maxwell® RSC reagent kits are specific for
the Maxwell® RSC or RSC 48 Instruments.
Related Products
Instrument and Accessories
Product | Size | Cat.# |
---|---|---|
Maxwell® RSC Instrument | 1 each | AS4500 |
Maxwell® RSC 48 Instrument | 1 each | AS8500 |
Maxwell® RSC/CSC Deck Tray | 1 each | SP6019 |
Maxwell® RSC/CSC 48 Front Deck Tray | 1 each | AS8401 |
Maxwell® RSC/CSC 48 Back Deck Tray | 1 each | AS8402 |
MaxprepTh Carrier, Maxwell® RSC | 1 each | AS9402 |
MaxprepTh Carrier, Maxwell® RSC 48 Front | 1 each | AS9403 |
MaxprepTh Carrier, Maxwell® RSC 48 Back | 1 each | AS9404 |
MaxprepTh Liquid Handler, RSC Carriers w/UV light | 1 each | AS9101 |
MaxprepTh Liquid Handler, RSC Carriers | 1 each | AS9100 |
MaxprepTh Liquid Handler, RSC 48 Carriers w/UV light | 1 each | AS9201 |
MaxprepTh Liquid Handler, RSC 48 Carriers | 1 each | AS9200 |
2.0ml Deep Well Plates (Non-Sterile) | 60/pack | AS9309 |
Nunc 2.0m1 Deep Well Plates (Sterile) | 60/pack | AS9307 |
MaxprepTh 1000p1 Conductive Disposable Tips, Filtered | 40/box | AS9303 |
MaxprepTh 300p1 Conductive Disposable Tips, Filtered | 60/box | AS9302 |
MaxprepTh Reagent Reservoir, 50m1 | 28/pack | AS9304 |
MaxprepTh Plunger Holder | 1 each | AS9408 |
MaxprepTh 3-Position Reagent Tube Holder | 1 each | AS9409 |
MaxprepTh Waste Bags, Clear | 100/box | AS9305 |
Maxwell® RSC Plunger Pack | 48/pack | AS1670 |
Maxwell® RSC/CSC Plungers | 50/pack | AS1331 |
RNase A Solution, 4ing/m1 | 1ml | A7973 |
ClickFlt Microhtbe, 1.5ml | 1,000/pack | V4741 |
Maxwell ® RSC Reagent Kits
Visit www.promega.com for a list of available
Maxwell ® RSC purification kits.
Summary of Changes
The following changes were made to the 3/22 revision of this document:
- Added new product Cat.# AS9309 to Sections 2 and 8.
- Updated product name for Cat.#AS9307 in Sections 2 and 8.
- Updated cover page.
(a) U.S. Pat. No. 6,855,499, European Pat. Nos. 1368629, 2090655 and 2363476,
Japanese Pat. No. 4399164 and other patents.
© 2014–2022 Promega Corporation. All Rights Reserved.
Maxwell is a registered trademark of Promega Corporation. MagnaCel and Maxprep
are trademarks of Promega Corporation.
Products may be covered by pending or issued patents or may have certain
limitations. Please visit our Web site for more information.
All prices and specifications are subject to change without prior notice.
Product claims are subject to change. Please contact Promega Technical
Services or access the Promega online catalog for the most up-to-date
information on Promega products.
Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TM419 · Revised 3/22
Documents / Resources
|
Promega AS1400 Maxwell RSC Blood DNA
Kit
[pdf] User Manual
AS1400, ASB1400, Maxwell RSC Blood DNA Kit
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