hygiena KIT230030 LP Plant Taxon Screening LyoKit User Guide
- June 16, 2024
- Hygiena
Table of Contents
KIT230030 LP Plant Taxon Screening LyoKit
foodproof®
Plant Taxon Screening LyoKit
Ready Reference Guide
Revision A, November 2023
Product No. KIT230030 (LP), KIT230031 (RP)
PCR kit for the qualitative detection of soya, maize, and rapeseed
(Brassicaceae) 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 (soya), HEX (maize), ROX (rapeseed) and Cy5 (Internal Control).
Pre-incubation: 1 cycle
Step 1: 37 °C for 4 min
Step 2: 95 °C for 10 min
Amplifi cation: 50 cycles
Step 1 : 95 °C for 5 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. 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 controls (H2 O),
before interpreting sample results.
Always compare samples to positive and negative control. Review data from each
channel and interpret results:
If a signal is present in channel _ sample is positive for respective target.
Examples for data interpretation:
| FAM | HEX | ROX | Cy5 | Result Interpretation |
|---|---|---|---|---|
| + | – | – | + or – | Positive for soya |
| – | + | – | + or – | Positive for maize |
| – | – | + | + or – | Positive for rapeseed |
| + | – | + | + or – | Positive for soya and rapeseed |
| + | + | + | + or – | Positive for all targets |
| – | – | – | + | Negative for all targets |
| – | – | – | – | Invalid |
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 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 fi lling, 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®
Plant Taxon Screening LyoKit
KIT230030 /31
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
Read User Manual Online (PDF format)
Read User Manual Online (PDF format) >>