QT-EVE-ZM-022
|
View in GMO-Matrix | View in GMO-Amplicons |
General information
| Tested in: |
GMO event MIR162 | SYN-IR162-4 (Maize)
|
| Genetic target: | MIR162 insert |
| Description: | Quantitative PCR method for detection of maize event MIR162 (EURL GMFF, 2011) |
| Comments: | This method was also verified in the context of the maize GMO stack Bt11 x MIR162 x TC1507 x NK603 (EURL-VL-02/24) in a bridging study, applying the relative quantification with a two-standard curve approach instead of the originally validated ∆Cq approach and using Rox as passive reference instead of sulforhodamine. |
Method details
| Specificity: | Event specific |
| Analysis type: | Quantitative |
| Assay: | Real-time PCR (Simplex) |
| Amplicon sequence: |
GCGCGGTGTCATCTATGTTACTAGNNNNNNNNNTCTAGACAATTCAGTACATTAAAAACGTCCGCCANNNTCTGAAGGCA ACAGATAAGGCA |
| Amplicon size: | 92 bp |
| Oligos: |
| Type | Name(s) | Sequence | Length | Target |
|---|---|---|---|---|
| Primer forward | MIR162-f1 / | GCGCGGTGTCATCTATGTTACTAG | 24 bp | insert |
| Primer reverse | MIR162-r1 / | TGCCTTATCTGTTGCCTTCAGA | 22 bp | 3'-host genome |
| Probe | MIR162-p1 / | FAM-TCTAGACAATTCAGTACATTAAAAACGTCCGCCA-TAMRA | 34 bp |
