Detection method details
Name: |
QT-TAX-GH-021 |
Description: |
Quantitative real-time PCR (TaqMan) method for detection of putative cotton SAH7 protein gene (115bp) (EU-RL GMFF GMOMETHODS database as January 22, 2014).
Estimated number of target copies per haploid genome (Jacchia et al. 2018): 2 |
Comment: |
Target is the endogenous putative Sinapis Arabidopsis Homolog 7 (SAH7) protein gene of Gossypium hirsutum
Copies of the SAH7 protein gene are coded on the A-subgenome and the D-subgenome of Gossypium hirsutum. Both are detected with the method QT-TAX-GH-021 but the respective amplicons differ in length (A-subgenome: 115 bp, D-subgenome: 123 bp). The assay for AdhC should be preferred for GMO quantitative analysis, but SAH7 can also be used because primers and probe anneal without mismatches to both target DNAs. |
Validation: |
ring trial validation |
Standardisation: |
EU reference method |
Related Methods: |
Type: |
taxon-specific |
Target Species name: |
Gossypium hirsutum |
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Oligonucleotides:
Forward Primer
Name:
Sah7-uni-f1Sequence:
AGTTTGTAGGTTTTGATGTTACATTGAGSize:
28
Reverse Primer
Name:
Sah7-uni-r1Sequence:
GCATCTTTGAACCGCCTACTGSize:
21
Probe
Name:
Sah7-uni-s1Sequence:
AAACATAAAATAATGGGAACAACCATGACATGTSize:
33
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Amplicon:
Sequence:
agtttgtaggttttgatgttacattgagnnnnnnnnnnnnnnnnnnnnnnnaaacataaaataatgggaacaaccatgacatgtnnnnnnnnnncagtaggcggttcaaagatgcSize:
115
Documents
Citation | Type | Local copy |
---|---|---|
Jacchia S., Kagkli D.F., Lievens A., Angers-Loustau A., Savini C., Emons H., Mazzara M. (2018) Identification of single target taxon-specific reference assays for the most commonly genetically transformed crops using digital droplet PCR. Food Control 93,191-200 | peer-reviewed article | |
EU-RL GMFF GMOMETHODS database; QT-TAX-GH-021 | EU reference method protocol |