hygiena KIT230053 GMO RR 2 Yield Soya Quantification Kit Instruction Manual
- June 17, 2024
- Hygiena
Table of Contents
KIT230053 GMO RR 2 Yield Soya Quantification Kit
foodproof®
GMO RR 2 Yield Soya Quantification Kit
Ready Reference Guide
Revision A, December 2023
Product No. KIT230053
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: MON89788 and Soya (Le1)
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 s
Step 2*: 60 °C for 60 s
- Fluorescence detection
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.
PREPARATION OF STANDARD CURVE
Use Calibrator DNA (purple cap) and Dilution Buffer (blue cap) to prepare
dilutions according to table below.
For each dilution step, pipet 30 µL (60 µL for duplicates) Dilution Buffer
into a new reaction tube. Transfer 10 µL
(20 µL for duplicates) from preceding step to new dilution step. Mix well
between pipetting steps.
The prepared dilutions can be used for both standard curves, GMO gene and
reference gene.
A typical experiment consists of 16 wells needed for standards and controls,
plus 2 × n wells (n = number of food samples). Since a multiwell plate has 96
wells, 40 food samples can be analyzed during one PCR run if the GMO gene and
the reference gene are analyzed in the same run. Some real-time PCR
instruments provide the opportunity to import external standard curves
generated in a previous run, then 46 food samples can be analyzed during one
PCR run.
| Dilution Step | Dilution Factor | Final Concentration |
|---|---|---|
| 1 | Undiluted | 100 |
| 2 | 1:04 | 25 |
| 3 | 1:16 | 6.25 |
| 4 | 0.086111 | 1.56 |
| 5 | 0.219444 | 0.39 |
| 6 | 0.752778 | 0.098 |
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 briefly spin vials before opening.
For data interpretation and calculation please refer to the entire product
manual.
-
ADD PCR MIX
For each master mix – the GMO Gene (yellow cap) and the Reference Gene (green cap) – pipet 20 µL into different strip or plate wells for the number of samples, standards and negative control.
-
ADD SAMPLES AND CONTROLS
Sample DNA must be diluted at least 1:4 in the Dilution Buffer (blue cap).
To each PCR mix (GMO and Reference), pipet 5 µL of samples, standards, negative control (colorless cap) or Control Template (purple cap) into respective wells.
-
SEAL
Seal strips/plate accurately.
-
CENTRIFUGE
Briefl y spin strips/plate in a suitable centrifuge.
-
START REAL-TIME PCR RUN
Cycle samples as described above.
foodproof®
GMO RR 2 Yield Soya
Quantification Kit
KIT230053
Kit for 2x 64 reactions
Store kit at -15 to -25 °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) >>