Table 3.
Real-time qPCR amplification of DNA from SU canola event 40K, Clearfield canola variety 5545 CL and 20 varieties of wild-type canola with PCR systems specific for the CruA endogenous canola reference gene and the AHAS1C-SU SNV present in SU canola. PCR conditions are as described in Methods, with 300 ng input DNA per reaction for each variety except that the concentrations of DNA in the reactions labeled 40K-10, 40K-1 and 40K-0.1 were 300, 30 and 3 ng DNA of 40K DNA, respectively, with total DNA adjusted to 300 ng with DNA from canola variety 5454 CL for reactions 40K-1 and 40K-01. For all wild-type varieties, n = 4. ND indicates that no amplification was detected; NA indicates “Not Applicable” because no amplification was observed.
| PCR Specificity | AHAS1C-SU | CruA | ||
|---|---|---|---|---|
| Canola Variety | Mean Ct | Ct% CV | Mean Ct | Ct% CV |
| 40K-10 | 25.36 | 0.08% | 19.88 | 0.20% |
| 40K-1 | 28.19 | 2.09% | 20.10 | 0.72% |
| 40K-0.1 | 33.43 | 1.44% | 20.22 | 0.12% |
| 5545 CL | ND | NA | 20.88 | 0.46% |
| Variety 1 | ND | NA | 21.25 | 0.20% |
| Variety 2 | ND | NA | 19.64 | 0.75% |
| Variety 3 | ND | NA | 20.15 | 0.35% |
| Variety 4 | ND | NA | 19.45 | 0.11% |
| Variety 5 | ND | NA | 19.37 | 0.52% |
| Variety 6 | ND | NA | 18.74 | 0.49% |
| Variety 7 | ND | NA | 19.27 | 0.19% |
| Variety 8 | ND | NA | 19.19 | 0.48% |
| Variety 9 | ND | NA | 19.22 | 0.55% |
| Variety 10 | ND | NA | 19.67 | 0.73% |
| Variety 11 | ND | NA | 19.56 | 0.78% |
| Variety 12 | ND | NA | 19.12 | 0.38% |
| Variety 13 | ND | NA | 19.45 | 0.19% |
| Variety 14 | ND | NA | 19.08 | 0.49% |
| Variety 15 | ND | NA | 19.62 | 0.28% |
| Variety 16 | ND | NA | 20.47 | 1.12% |
| Variety 17 | ND | NA | 21.23 | 0.20% |
| Variety 18 | ND | NA | 19.06 | 0.50% |
| Variety 19 | ND | NA | 23.65 | 0.26% |
| Variety 20 | ND | NA | 18.29 | 0.39% |
| No DNA | ND | NA | 34.36 | 2.48% |