hygiena MON88017 GMO Maize Multiplex Detection Kit User Guide

hygiena MON88017 GMO Maize Multiplex Detection Kit

GMO Maize Multiplex Detection Kit

Specifications

• Product Name: GMO Maize Multiplex Detection Kit
• Product Number: KIT230219
• Revision: A
• Release Date: December 2023
• PCR Kit for the qualitative detection of MON88017, NK603, and MIR162 DNA
• For use with real-time PCR instruments
• Kit Size: 50 reactions
• Intended Use: Food testing purposes, for in vitro use only

Program Setup
Before setting up the PCR reactions, program your real-time PCR instrument as follows:

• Select the following channels: u FAM (MON88017), VIC/HEX (NK603), ROX (MIR162), and Cy5 (Internal Control)
• Set the cycling parameters as follows:
  • 2 minutes at 10°C
  • 10 minutes at 60°C
  • 1 cycle of 15 seconds at 95°C and 60 seconds at 60°C
  • 40 cycles of 15 seconds at 95°C and 60 seconds at 60°C (Fluorescence detection)
• Note: For some real-time PCR instruments, you may need to specify the probe quencher and use of a passive reference dye. This kit contains probes with a non-fluorescent dark quencher and no passive reference dye.

Data Interpretation
Before interpreting the sample results, verify the results of the positive (Control Template) and negative (H2O) controls. Always compare the samples to the positive and negative controls. Review the data from each channel and interpret the results as described in the table below:

Preparation of the PCR Mix
Before starting, take appropriate precautions to prevent contamination, such as using filter tips and wearing gloves. Follow the steps below to prepare the PCR mix:

1. Thaw the reagents and briefly spin the vials before opening (do not vortex!)
2. Prepare the PCR mix for each reaction in a suitable tube
   • The number of reactions should be equal to the number of samples plus 2 controls, plus at least one additional reaction to cover pipetting loss
3. Mix the PCR mix carefully but thoroughly by pipetting up and down
4. Add the PCR mix to each reaction
5. Add the samples and controls to the respective wells
6. Carefully seal the strips/plate
7. Briefly spin the strips/plate in a suitable centrifuge
8. Start the real-time PCR run using the cycling parameters mentioned in the program setup section

FAQ (Frequently Asked Questions)

• Q: Can this kit be used for human DNA testing?
  A: No, this kit is specifically designed for the qualitative detection of MON88017, NK603, and MIR162 DNA in maize samples for food testing purposes. It is not intended for human DNA testing.

• Q: Can I use this kit with any real-time PCR instrument?
  A: The kit is compatible with most real-time PCR instruments. However, for some instruments, you may need to specify the probe quencher and use of a passive reference dye. Please refer to your instrument’s manual or contact our customer support for compatibility information.

foodproof® SL
GMO Maize Multiplex Detection Kit (MON88017, NK603, MIR162)

Ready Reference Guide
Revision A, December 2023

Product No. KIT230219
PCR kit for the qualitative detection of MON88017, NK603 and MIR162 DNA using real-time PCR instruments. Before starting, it is strongly recommended to read the entire product manual available on our website.

PROGRAM SETUP

• Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:
  • FAM (MON88017), VIC/HEX (NK603), ROX (MIR162) and Cy5 (Internal Control).

• Pre-incubation: 1 cycle
  • Step 1: 50 °C for 2 min
  • Step 2: 95 °C for 10 min
• Amplification: 40 cycles
  • Step 1 : 95 °C for 15 sec
  • Step 2*: 60 °C for 60 sec

* Fluorescence detection

For some real-time PCR instruments the probe quencher as well as the use of a passive reference dye must be specified. This kit contains probes with a non- fluorescent “dark” quencher and no passive reference dye.

DATA INTERPRETATION

Verify results of positive (Control Template) and negative (H2O) controls, before interpreting the sample results. Always compare samples to positive and negative controls. Review data from each channel and interpret results as described in the table.

PREPARATION OF THE PCR MIX

Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves. Thaw reagents, mix (do not vortex!) and briefly spin vials before opening.

1. PREPARE PCR MIX
   (n samples + 2 controls + at least one additional reaction to cover pipetting loss). Mix carefully but thoroughly by pipetting up and down.

2. ADD PCR MIX
   Pipette 20 µL of prepared PCR mix into each strip or plate well.

3. ADD SAMPLES AND CONTROLS
   Pipette 5 µL of samples, negative control (PCR-grade H2O) or Control Template (C) into respective wells.

4. SEAL
   Carefully seal strips/plate.

5. CENTRIFUGE
   Briefly spin strips/plate in a suitable centrifuge.

6. START REAL-TIME PCR RUN
   Cycle samples as described above.

foodproof® SL GMO

• Maize Multiplex Detection Kit (MON88017, NK603, MIR162)
• KIT230219
• Kit for 50 reactions Store kit at -15 to -25 °C
• For food testing purposes
• FOR IN VITRO USE ONLY

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

References

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