Processing
PrimeTime® Assays with Fast-Cycling Protocols—Comparative Data
| WDR3 | TIMP3 | HERC1 | ||
|---|---|---|---|---|
| Mean Cq (50 ng) | PrimeTime® | 22.9 | 23.9 | 25.1 |
| Competitor A | 24.0 | 24.1 | 26.2 | |
| Efficiency | PrimeTime® | 97.0% | 95.0% | 95.9% |
| Competitor A | 96.4% | 91.7% | 86.8% | |
| Endpoint (ΔRn) | PrimeTime® | 10.9 | 8.9 | 7.9 |
| Competitor A | 7.2 | 6.1 | 7.0 | |
PrimeTime® qPCR Assays Maintain Efficiency Under Fast-Cycling Conditions. 25 PrimeTime qPCR Assays (blue curves) were compared to equivalent Competitor A assays (consisting of 15 inventoried pre-designed assays and 10 made-to-order pre-designed assays; red curves). To ensure an accurate comparison was made, the PrimeTime Assays and Competitor A assays were selected to span the same exon boundary of each gene. Reactions included five 4-fold dilutions cDNA template and the Applied Biosystems TaqMan® Gene Expression Master Mix, and were run on the ABI 7900HT Fast Real-Time PCR System with the following PCR cycling conditions: 2 min. 50°C; 10 min. 95°C; 45 x (15 sec. 95°C, 1 min. 60°C). Identical thresholds were set for all runs for comparison across assays.