PrimeTime® Assays with Fast-Cycling Protocols—Comparative Data

Mean Cq (50 ng)PrimeTime®22.923.925.1
Competitor A24.024.126.2
Competitor A96.4%91.7%86.8%
Endpoint (ΔRn)PrimeTime®
Competitor A7.26.17.0

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.