hygiena KIT230022 LP GMO Maize Identification 1 LyoKit User Guide

hygiena KIT230022 LP GMO Maize Identification 1 LyoKit

Specifications:

• Product No.: KIT230022 (LP), KIT230023 (RP)
• Revision: A, November 2023
• PCR kit for the qualitative detection of DAS-40278-9, LY038, and VCO-01981-5 DNA using real-time PCR instruments

Program Setup:

Before setting up the PCR reactions, program your real-time PCR instrument with the following settings:

• FAM (DAS-40278-9) – Fluorescence detection
• HEX (LY038) – Fluorescence detection
• ROX (VCO-01981-5) – Fluorescence detection

Please ensure that you have selected the correct channels for each target DNA.

Data Interpretation:

To interpret the sample results, follow these steps:

1. Verify results of positive (Control Template) and negative (H2O) controls.
2. Compare samples to positive and negative controls.
3. Review data from each channel.
4. Interpret results based on the presence or absence of a signal in each channel.

The following examples demonstrate result interpretation:

Result Interpretation:

• Positive for DAS-40278-9
• Positive for LY038
• Positive for VCO-01981-5
• Negative for all targets

Preparation of the PCR Mix:

Follow these steps to prepare the PCR mix:

1. Place Strips in Rack: Take the required number of PCR tube strips out of the aluminum bag. Close the bag tightly afterwards. Place the strips in a suitable PCR tube rack. Gently tap the tubes, if needed, to move the lyophilized pellets to the bottom of all tubes.
2. Decap: Immediately before filling, carefully open the strips and discard the caps. Do not leave them open longer than necessary.
3. Add Samples and Controls: Add the samples and controls to the tubes.
4. Seal: Carefully seal the tubes with the provided 8-cap strips.
5. Mix: Resuspend the pellet after sealing by mixing thoroughly. Alternatively, resuspend the pellet by pipetting up and down multiple times in Step 3.
6. Centrifuge: Briefly spin the strips, e.g., 5 seconds at 500 – 1,000 x g, in a suitable centrifuge.
7. Start Real-Time PCR Run: Cycle the samples as described in the program setup section. Place the tubes in a vertical, balanced order into the cycler.

For food testing purposes FOR IN VITRO USE ONLY Made in Germany

FAQ:

Q: Can I reuse the PCR tube strips?

A: No, the PCR tube strips are for single-use only. Discard them after each use to prevent contamination.

Q: What precautions should I take to prevent contamination?

A: To prevent contamination, use filter tips and wear gloves when handling the PCR mix and samples.

Q: Can I use this kit for GMO maize identification in other crops?

A: No, this kit is specifically designed for the qualitative detection of DAS-40278-9, LY038, and VCO-01981-5 DNA in GMO maize samples.

Revision A, November 2023

Product No. KIT230022 (LP), KIT230023 (RP)

PCR kit for the qualitative detection of DAS-40278-9, LY038 and VCO-01981-5 DNA using real-time PCR instruments.

PROGRAM SETUP

Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:

• FAM (DAS-40278-9), HEX (LY038) and ROX (VCO-01981-5).

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.
A Color Compensation is necessary for users of the LightCycler® 480 System: Color Compensation Set 3 (Product No. KIT230005).

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:
If a signal is present in channel _ sample is positive for the respective target.
Examples for data interpretation:

PREPARATION OF THE PCR MIX

Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves.

1. PLACE STRIPS IN RACK
   Take needed number of PCR tube strips out of aluminum bag.
   Important: close bag tightly afterwards. Place strips in a suitable PCR tube rack.
   If needed, gently tap the tubes to move the lyophilized pellets to the bottom of all tubes.

2. DECAP
   Immediately before filling, carefully open strips and discard caps.
   Do not leave open longer than necessary.

3. ADD SAMPLES AND CONTROLS
   Pipette 25 µL of samples, Negative Control (colorless cap) or Control Template (purple cap) into respective wells.
   If using less volume, add PCR-grade H2O to reach 25 µL.

4. SEAL
   Carefully seal the tubes with the provided 8-cap strips.

5. MIX
   Resuspend pellet after sealing by mixing thoroughly.
   Alternatively, resuspend pellet by pipetting up and down multiple times in Step 3.

6. CENTRIFUGE
   Briefly spin strips, e.g., 5 seconds at 500 – 1,000 x g, in a suitable centrifuge.

7. START REAL-TIME PCR RUN
   Cycle samples as described above.
   Place tubes in a vertical, balanced order into the cycler, e.g., two strips can be placed in the first and last column.

foodproof® GMO Maize Identification 1 LyoKit

KIT230022 /23
Kit for 96 reactions Store kit at 2 to 8 °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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