hygiena KIT230090 Plant Detection Lyo Kit User Guide

hygiena KIT230090 Plant Detection Lyo Kit

Specifications:

• Product No.: KIT230090 (LP), KIT230091 (RP)
• Revision: A, November 2023
• PCR kit for the qualitative detection of Plant DNA
• Compatible with real-time PCR instruments
• Contains probes with a non-fluorescent dark quencher and no passive reference dye

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

• FAM (Plant) channel
• HEX (Internal Control) channel

The program setup should include the following cycle:

• 4 minutes at an initial temperature
• 10 minutes at a second temperature
• 1 cycle of 15 seconds and 60 seconds for fluorescence detection
• 50 cycles of PCR amplification

Data Interpretation:
After running the PCR reactions, verify the results by comparing them to the positive control (Control Template) and negative control (H2O). The interpretation of the results is as follows:

| FAM| HEX
---|---|---
Result Interpretation| Positive for Plant| Negative for Plant| Invalid

Preparation of the PCR Mix:
Follow these steps to prepare the PCR mix:

1. Place the needed number of PCR tube strips out of the aluminum bag and close the bag tightly.
2. Carefully open the strips and discard the caps.
3. Add samples and controls to the open tubes.
4. Seal the tubes with the provided 8-cap strips.
5. Mix the contents by resuspending the pellet thoroughly. Alternatively, resuspend the pellet by pipetting up and down multiple times in Step 3.
6. Briefly spin the strips in a suitable centrifuge (e.g., 5 seconds at 500 – 1,000 x g).
7. Start the real-time PCR run using the programmed settings.

FAQ:

• Q: What channels should I select on my real-time PCR instrument?
  A: Select the FAM (Plant) and HEX (Internal Control) channels.

• Q: Can I use this kit with any real-time PCR instrument?
  A: This kit is compatible with most real-time PCR instruments. However, for some instruments, you may need to specify the probe quencher and usage of a passive reference dye.

• Q: How should I interpret the PCR results?
  A: Always compare your samples to the positive and negative controls. Review the data from each channel (FAM and HEX) and  interpret the results according to the provided table.

• Q: How should I prevent contamination during preparation?
  A: Take appropriate precautions, such as using filter tips and wearing gloves, to prevent contamination.

• Q: Can I leave the PCR tube strips open for an extended period?
  A: No, it is recommended to not leave the strips open longer than necessary to minimize the risk of contamination.

• Q: How should I place the tubes in the real-time PCR instrument?
  A: Place the tubes in a vertical, balanced order into the cycler. For example, two strips can be placed in the first and last column.

• Q: Can I use this kit for food testing purposes?
  A: Yes, this kit can be used for food testing purposes. However, it is for in vitro use only.

• Q: Where is this kit manufactured?
  A: This kit is made in Germany.

Plant Detection LyoKit Ready Reference Guide
Product No. KIT230090 (LP), KIT230091 (RP)

PCR kit for the qualitative detection of Plant 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 (Plant) and HEX (Internal Control).

For some real-time PCR instruments, the probe quencher as well as the usage of a passive reference dye has to 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 controls (H2O), before interpreting sample results. Always compare samples to positive and negative control. 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 fi lter tips and wearing gloves.

1. PLACE STRIPS IN THE RACK
   Take the needed number of PCR tube strips out of an aluminum bag.
   Important : close the bag tightly afterward. 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 it 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 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 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®
Plant Detection LyoKit
KIT230090 / 91
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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