PCRmax Genesig FMV 35S promoter in GM crops Instruction Manual
FMV 35S promoter
FMV 35S promoter
(GMO)
PCRmax Ltd qPCR test
TM
150 tests
For general laboratory and research use only
1
Quantification of FMV 35S promoter (GMO) genomes.
Advanced kit handbook HB10.07.08
Published Date: 26/04/2016
The kit provides a method for detecting gene insertion events by real-time PCR. The kit is
based on the PCR amplification and detection of the Figwort mosaic virus (FMV) promoter.
This promoter is preferred above other potential promoters because it is highly
transcriptionally active and is not greatly influenced by environmental conditions or tissue
types.
Since FMV is naturally found in the soil, it is necessary to exclude false positive results by
proving that the wild type virus has not infected or contaminated a non GM sample. This
kit
provides a control test that detects a region of the FMV genome that is not used in the
genetic modification of plants.
A positive signal with the FMV-GM test indicates the presence of GM material in the
sample, provided that no signal is obtained from the FMV-WT test.
Where signals are obtained with the FMV-WT test this indicates that naturally occurring
FMV is present in the samples. The data must be carefully considered by looking at the
relative signal strength between the FMV-WT test and the FMV-GM test in the samples.
Within the details result view you can review the Cq values for each of the tests. If the
FMV-GM test detection occurs 5 or more Cq values earlier than the FMV-WT control test
then the test confirms the presence of both the wild FMV organism and GM plant material.
If the signal strength for the FMV-WT and FMV-GM are closer together with a
smaller difference in Cq values then the test is indeterminate since the presence of wild
FMV prevents the analysis of the GM content of the sample.
Introduction to FMV 35S promoter (GMO)
2
Quantification of FMV 35S promoter (GMO) genomes.
Advanced kit handbook HB10.07.08
Published Date: 26/04/2016
MAX
MIN
This kit is designed to have 100% homology with all available sequence data on the
Figwort mosaic virus (FMV) promoter.
The control test detects a region of the FMV genome that is not used in the genetic
modification of plants.
If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to help@pcrmax.com and our bioinformatics team will
answer your question.
Specificity
3
Quantification of FMV 35S promoter (GMO) genomes.
Advanced kit handbook HB10.07.08
Published Date: 26/04/2016
Kit Contents
• FMV 35S promoter primer/probe mix (150 reactions BROWN)
FAM labelled
• FMV Wild-type virus primer/probe mix (150 reactions BROWN)
FAM labelled
• FMV 35S promoter positive control template (for Standard curve RED)
• FMV Wild-type virus positive control template (for Standard curve RED)
• Internal extraction control primer/probe mix (150 reactions BROWN)
VIC labelled as standard
• Internal extraction control DNA (150 reactions BLUE)
• Endogenous control primer/probe mix (150 reactions BROWN)
FAM labelled
• RNAse/DNAse free water (WHITE)
for resuspension of primer/probe mixes and internal extraction control DNA
• Template preparation buffer (YELLOW)
for resuspension of positive control templates and standard curve preparation
Reagents and equipment to be supplied by the user
Real-Time PCR Instrument
DNA extraction kit
This kit is designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.
Lyophilised 2x qPCR Mastermix
This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.
Pipettors and Tips
Vortex and centrifuge
Thin walled 1.5 ml PCR reaction tubes
4
Quantification of FMV 35S promoter (GMO) genomes.
Advanced kit handbook HB10.07.08
Published Date: 26/04/2016
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