PerfeCTa® qPCR Supermixes for more robust, convenient and
economic genotyping

PerfeCTa® qPCR Supermixes raise the bar for performance and economy by generating accurate allele calls while requiring less probe and primer. Greater productivity is achieved through faster run times, ease of use and stability.

Features & Benefits

• Shorter Run Times – enabling more experiments per day
• Robust Signal – use ½ the primer and probe concentration
• Convenience & Reliability – stability for 3 days pre-PCR and 5 days post-PCR
• Simplified & Robust Allele Calling – fewer sample failures or re-work

Description

PerfeCTa® qPCR SuperMixes are 2X concentrated, ready-to-use reaction cocktails containing all components except primers, probe and template for any real-time quantitative PCR instrument. AccuStart™ Taq enables faster activation and extension rate resulting in enhanced productivity through faster cycling (see table comparison). Superior signal generation of PerfeCTa® SuperMixes allows greater economy through use of lower primer and probe concentrations compared to other leading products (see Fig. 1). Additionally PerfeCTa® SuperMixes yield reliable discrimination of SNPs in difficult targets (see Fig. 2).

Fig. 1 Speed. Obtain equivalent SNP results with ½ primer-probe in ½ the time. Genotyping reactions were performed on a panel of 94 gDNA samples (3 ng) and 2 NTCs with either TaqMan® Genotyping Master Mix and 1X TaqMan® SNP Assay C__12050942_10 or PerfeCTa™ qPCR SuperMix with 1/2X primer/probe mix. Cycling protocols were as described in Table 1 followed by allelic discrimination analysis on the 7500 System (Applied Biosystems).

Fig. 2. Signal-to-noise ratio. Obtain higher and discrete fluorescent signals for more accurate allele calls. Comparison of TaqMan® Genotyping Master Mix and PerfeCTa™ qPCR SuperMix for allelic discrimination of an A/G transition in CYP2C19. Genotyping reactions were run on a panel of 94 gDNA samples (3 ng) and two NTCs according to the each manufacture’s recommend conditions with TaqMan® SNP Assay C__25986767_70 (Applied Biosystems).

Comparison Conventional vs. PerfeCTa®

Quanta SuperMixes include optimized buffers and stabilizers for TaqMan® probe and MGB-probe chemistries. The enhanced specificity of PerfeCTa® SuperMixes suppress cross-reactivity between homologous sequences, improving detection and discrimination in SNP applications. This provides discrete and well-separated clusters for clear genotyping results. Tight control of AccuStart™ Taq polymerase activity conveys exceptional pre-PCR stability for convenient room temperature reaction assembly and flexible scheduling of PCR runs (see Fig. 3). Stable fluorescent signal allows automated scanning of results without compromising assay performance (see Fig. 4).

Fig. 3 Pre-PCR Stability Tight control of Taq polymerase activity in PerfeCTa™ qPCR SuperMix provides excellent Pre-PCR reaction stability for flexible scheduling of automated PCR runs. Replicate 96-well D11S13634351376 genotyping reactions with PerfeCTa™ qPCR SuperMix were performed as described in Fig 1. Plate A (blue) was cycled immediately, plate B (red) was held at room temperature for 72 hrs before PCR amplification.

Fig. 4 Post-PCR Stability Proprietary fluorescence stabilizers in PerfeCTa™ qPCR SuperMix facilitate automated data acquisition and consistent genotyping results. 7500 System allelic discrimination analysis of plate A from the preceding figure was performed either immediately following PCR (blue) or after sitting on the bench for 5 days (red).

Product List Genotyping