VeriFi Hot Start Polymerase & Mixs

AptaLock™ is an innovative proprietary technology that uses an aptamer-like molecule to confer hot start activity to the polymerase. It is an alternative method to an antibody-mediated hot start.  The AptaLock™ molecule reversibly binds and inhibits the exonuclease and polymerase activities of the enzyme at temperatures below 40°C. This allows the reaction to be set up at room temperature without the loss of sensitivity and specificity as a result of primer dimer formation and non-specific amplification.

Figure 1. Superior performance in multiplex reactions
10-plex PCR using lambda phage genome (6 targets) and mouse genome (4 targets) at different annealing temperatures (A: 63.0°C, B: 61.5°C, C: 60.5°C). The starting template amount is 1pg lambda DNA and 1ng mouse gDNA. Amplicon lengths are between 139bp and 962bp. Reactions were set up using master mix formats following manufacturers’ recommendations. Cycling conditions were 95°C 2 min, 40 cycles of 95°C 15 sec, annealing A to C 30 sec, 72°C 90 sec. L:
PCRBIO Ladder III. P: reference pool of single products. PCRBIO HS VeriFi™ Mix displays greater sensitivity and specificity in multiplex when compared to leading competitors

Figure 2. Successful PCR across a broad range of GC and AT content
Amplification of 13 targets with GC content ranging from 28.7% to 83% using PCRBIO HS VeriFi™ Mix. The starting template amount is 30ng mouse cDNA. Band size is between 99bp and 274bp. Cycling conditions were 98°C 5 min, 40 cycles of 98°C 15 sec, annealing between 54°C and 62°C (depending on target) 15 sec, 72°C 30 sec. L: PCRBIO Ladder III. PCRBIO HS VeriFi™ Mix is able to amplify templates across a broad range of GC and AT content.

Figure 3. Increased PCR success rates and consistency with complex targets
Amplification of a 13.5kb fragment of the human ß-globin gene at different annealing temperatures (A: 68.5°C, B: 66.0°C, C: 63.0°C, D: 60.5°C). The starting template amount is 30ng human genomic DNA. GC content is 37%. Reactions were set up using master mix formats (apart from Takara’s PrimeSTAR® GXL DNA Polymerase) and following manufacturers’ recommendations. Cycling conditions were 95°C for 2 min, then 30 cycles of 95°C for 15 sec, annealing for 15 sec, 72°C for 12 min. PCRBIO HS VeriFi™ Mix displays a higher yield and specificity compared to leading competitors. The PCRBIO mix also shows greater consistency and versatility across the annealing temperature range.