hygiena KIT230108 LP GMO Soya Identification 3 LyoKit User Guide
- June 16, 2024
- Hygiena
Table of Contents
hygiena KIT230108 LP GMO Soya Identification 3 LyoKit User Guide
Ready Reference Guide
Product No. KIT230108 (LP), KIT230109 (RP)
PCR kit for the qualitative detection of FG72, A5547-127, DP-356043 and
MON87705 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:
u DAM (FG72), HEX (A5547-127), XOR (DP-356043) and Cyst (MON87705
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
1 cycle * 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 (H2 O) 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 the channel _ the sample is positive for the
respective target.
Examples for data interpretation:
| FAM | HEX | ROX | Cy5 | Result Interpretation |
|---|---|---|---|---|
| + | – | – | – | Positive for FG72 |
| – | + | – | – | Positive for A5547-127 |
| – | – | + | – | Positive for DP-356043 |
| – | – | – | + | Positive for MON87705 |
| + | – | + | – | Positive for FG72 and DP-356043 |
| + | + | + | – | Positive for FG72, A5547-127 and DP-356043 |
| – | – | – | – | Negative for all targets |
PREPARATION OF THE PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using fi lter tips and wearing gloves.
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.
DECAP
Immediately before fi lling, carefully open strips and discard caps. Do not
leave open longer than necessary
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 H2 O
to reach 25 µL
SEAL
Carefully seal the tubes with the provided 8-cap strips.
MIX
Resuspend pellet after sealing by mixing thoroughly. Alternatively, resuspend
pellet by pipetting up and down multiple times in Step 3.
CENTRIFUGE
Briefl y spin strips, e.g., 5 seconds at 500 – 1,000 x g, in a suitable
centrifuge.
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 fi rst and last column.
foolproof® GMO Sonya
Identification 3 Toolkit
KIT230108 / 09
Kit for 96 reactions
Store kit at 2 to 8 °C
For food testing purposes
FOR IN NITRO USE ONLY
Made in Germany
Hygiena® | Camarillo, CA 93012 USA | diagnostics.support@hygienically | hygienically