hygiena T25 GMO Maize Multiplex Detection Kit User Guide
- June 16, 2024
- Hygiena
Table of Contents
hygiena T25 GMO Maize Multiplex Detection Kit User Guide
PCR kit for the qualitative detection of T25, MON810 and MON863 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 (T25), VIC/HEX (MON810), ROX (MON863) and Cy5 (Internal Control).
Pre-incubation: 1 cycle
Step 1: 50 °C for 2 min
Step 2: 95 °C for 10 min
Amplifi cation: 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 (H2 O) 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.
| FAM | VIC/HEX | ROX | Cy5 | Result Interpretation |
|---|---|---|---|---|
| + | + | + | + or – | Positive for T25, MON810 and MON863 |
| – | + | + | + or – | Positive for MON810 and MON863 |
| + | – | + | + or – | Positive for T25 and MON863 |
| + | + | – | + or – | Positive for T25 and MON810 |
| – | + | – | + or – | Positive for MON810 |
| + | – | – | + or – | Positive for T25 |
| – | – | + | + or – | Positive for MON863 |
| – | – | – | + | Negative for T25, MON810 and MON863 |
| – | – | – | – | Invalid |
foodproof® is a registered trademark of Hygiena. The above mentioned real-time PCR instruments are registered trademarks of their respective holders.
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.
PREPARE PCR MIX
Add 12.5 µL Master Mix (2XM),
4.0 µL Primer/Probe Mix (P) and
3.5 µL PCR-grade H2 O (not included) for each reaction toa suitable tube
(n samples + 2 controls + at least one additional reaction to cover pipetting loss). Mix carefully but thoroughly by pipetting up and down.
ADD PCR MIX
Pipette 20 µL of prepared PCR mix into each strip or plate well.
ADD SAMPLES AND CONTROLS
Pipette 5 µL of samples, negative control (PCR-grade H2 O) or Control Template (C) into respective wells.
SEAL
Carefully seal strips/plate.
CENTRIFUGE
Briefl y spin strips/plate in a suitable centrifuge.
START REAL-TIME PCR RUN
Cycle samples as described above.
foodproof® SL GMO Maize
Multiplex Detection Kit
(T25, MON810, MON863)
KIT230221
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