hygiena Bt11 GMO Maize Multiplex Detection Kit User Guide
- June 16, 2024
- Hygiena
Table of Contents
food proof® SL
GMO Maize Multiplex Detection Kit (Bt11, TC1507)
Ready Reference Guide
Revision A, December 2023
Product No. KIT230220
PCR kit for the qualitative detection of Bt11 and TC1507 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 (Bt11), VIC/HEX (TC1507) and Cy5 (Internal Control).
- 1 cycle
- 40 cycles
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.
FAM| VIC/HEX|
Cy5
| Result Interpretation
---|---|---|---
| +|
+ or –
| Positive for Bt11 and TC1507
–
| +|
+ or –
| Positive for TC1507
| –|
+ or –
| Positive for Bt11
–
| –|
| Negative for Bt11 and TC1507
–
| –|
–
| Invalid
food proof® is a registered trademark of Hygiena. The above mentioned real-time PCR instruments are registered trademarks of their respective holders.
RR-KIT230220-RevA
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.
A: +12.5 µL + 4 µL + 3.5 µL
1. PREPARE PCR MIX
Add 12.5 µL Master Mix (2XM), 4.0 µL Primer/Probe Mix (P) and 3.5 µL PCR-
grade H2O (not included) → for each reaction to a suitable tube.
(n samples + 2 controls + at least one additional reaction to cover pipetting
loss).
Mix carefully but thoroughly by pipetting up and down.
A: +20 µL
2. ADD PCR MIX
Pipette 20 µL of prepared PCR mix into each strip or plate well.
A: +5 µL
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.
food proof® KIT230220 For food
testing purposes
SL GMO Maize Multiplex Kit for 50 reactions FOR IN VITRO USE ONLY
Detection Kit (Bt11, TC1507) Store kit at -15 to -25 °C
Hygiena® | Camarillo, CA 93012 USA | diagnostics.support@hygiena.com | www.hygiena.com