hygiena KIT230011 Color Compensation Set 5 Owner’s Manual

hygiena KIT230011 Color Compensation Set 5

Color Compensation Set 5 Ready Reference Guide

Specifications

• Product No.: KIT230011
• Revision: A, December 2023
• Color Compensation Set 5 Kit for 5 runs
• Store kit in the dark
• For food testing purposes
• FOR IN VITRO USE ONLY
• Made in Germany

Product Usage Instructions

Program Setup:
Ensure the following settings for the PCR program:

• Temperature Gradient: 2.2 –
• Color Compensation: 1
• Cooling: None
• Parameter Settings:
  • Name: FAM HEX ROX Cy5
  • Melt Factor: 1 1 1 1
  • Quant Factor: 10 10 10 10
  • Max Integration Time (s): 2 2 2 2

Preparation of the PCR Mix:

1. Add blank to the first well.
2. Add blank to additional wells.
3. Add calibrator dyes.
4. Seal & centrifuge: Seal strips accurately and briefly spin strips/plate in a suitable centrifuge.
5. Cycle samples:
   • Vials: 0 1 2 3 4
   • Dominant Channel: water 465 – 510 (FAM) 533 – 580 (HEX) 533 – 610 (ROX) 618 – 660 (Cy5)

Analysis:
Follow these steps for analysis:

1. In the Subset Editor, create a new subset for the Color Compensation reactions.
2. In the Sample Editor, define the dominant channel for each position.
3. Select Color Compensation from the analysis menu, click Calculate, and then click the Save CC Object button.

FAQ:

• Q: How should I store the Color Compensation Set 5 Kit?
  A: Store the kit in the dark to maintain its integrity.

• Q: Can this kit be used for purposes other than food testing?
  A: No, this kit is specifically designed for food testing purposes and should only be used for in vitro applications.

Color Compensation Set 5
Ready Reference Guide

• Product No. KIT230011
• Revision A, December 2023

Color compensation set for the generation of color compensation objects for the LightCycler® 480 System. The set is designed for 5 calibration runs.
Before starting, it is strongly recommended to read the entire product manual available on our website.

PROGRAM SETUP

Program the LightCycler® 480 instrument as shown below before preparing the calibration mixes.

| | |
---|---|---|---
| |
|
| | |
| | | |
| Temperature Gradient| Cooling
Cycles| 1| 1
Analysis Mode| Color Compensation| None
Steps| 1| 2| 3| 1
Target Temp| 95 °C| 37 °C| 95 °C| 40 °C
Acquisition Mode| None| None| Continuous| None
Hold| 1 s| 10 s| –| 30 s
Ramp Rate| 4.4| 2.2| –| 2.2
Acquisition/°C| –| –| 1| –
Parameter| Setting| Name| Melt Factor| Quant Factor| Max Integration Time (s)
---|---|---|---|---|---

Detection Format

| 465 – 510| FAM| 1| 10| 2
533 – 580| HEX| 1| 10| 2
533 – 610| ROX| 1| 10| 2
618 – 660| Cy5| 1| 10| 2
Block Size| 96|
Reaction Volume| 25 µl

PREPARATION OF THE PCR MIX

Take appropriate precautions to prevent contamination, e.g., by using fi lter tips and wearing gloves.
Thaw reagents, mix (do not vortex!), and briefl y spin vials before opening. Use tube strips and caps matching Hygiena® Diagnostics’ LyoKit consumables.

1. ADD BLANK TO FIRST WELL
   Pipet 25 µL of Blank (vial 0, white cap) into fi rst well.

2. ADD BLANK TO ADDITIONAL WELLS
   Pipet 20 µL of Blank (vial 0, white cap) into second, third, fourth and fi fth well.

3. ADD CALIBRATOR DYES
   Pipet 5 µL of Calibrator 1 (yellow cap), Calibrator 2 (red cap), Calibrator 3 (purple cap) and Calibrator 4 (blue cap) into second, third, fourth and fi fth well, respectively.

4. SEAL & CENTRIFUGE
   Seal strips accurately and briefl y spin strips/plate in a suitable centrifuge .

5. CYCLE SAMPLES
   Insert the PCR strips into a support frame matching LightCycler® plates and load to the instrument.
   Cycle the samples as described.

6. ANALYSIS
   In the Subset Editor, create a new subset for the Color Compensation reactions. In the Sample Editor, defi ne the dominant channel for each position.
   Select “Color Compensation” from the analysis menu, click “Calculate” and then click the “Save CC Object” button.

Color Compensation Set 5 KIT230011
Kit for 5 runs Store kit in the dark at -25 to -15 °C
For food testing purposes FOR IN VITRO USE ONLY Made in Germany

Hygiena® | Camarillo, CA 93012 USA | diagnostics.support@hygiena.com | www.hygiena.com

References

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