repliQa HiFi ToughMix
- High Fidelity – >90x wild type Taq
- Extreme Speed – up to 3x faster PCR results with extension rates as fast as 1 kb/sec*
- Tough Tested – tolerant to a wide range of PCR inhibitors
- Superior Sensitivity – higher yields from lower inputs as little as 2 pg
- Long Range – amplify +24 kb gDNA, +40 kb λDNA
*for fragments less than 1 kb in size
repliQa HiFi ToughMix is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
repliQa HiFi ToughMix
Description
The repliQa HiFi ToughMix is a 2x, ready-to-use master mix that contains all the components for high fidelity PCR amplification, including a genetically modified DNA polymerase coupled with hot start antibodies. This unique, next generation master mix provides 90x higher fidelity compared to Taq, while reducing time to PCR results by 2-3x. The extreme speed is enabled by extension times as fast as 1-10 kb/sec depending on target length. The enzyme is coupled with the industry leading ToughMix which is tolerant to a wide variety of inhibitors making it suitable for routine PCR, cloning, amplicon sequencing and site directed mutagenesis.
Details
2x reaction buffer containing optimized concentrations of MgCl2, dNTP’s and proprietarily formulated HiFi polymerase, hot start antibodies and ToughMix chemistry
Performance Data
Extreme Speed: Up to 3x faster results
Comparison of speed. A 2 kb fragment was amplified in 50 μl reaction volumes according to the recommended protocol. Following a 30 s activation at 98°C; 30 cycles of PCR were performed: 98°C, 10 s; 60°C, 10 s; 68°C, 5–30 s. The thermal cycler had a ramp rate of 5°C/s.
Tough Tested: Tolerant to a wide range of PCR inhibitors
Strong Inhibitor Resistance. A 2 kb λ DNA template was amplified using each manufacturer’s recommended cycling conditions with different amounts io inhibitors. This experiment was run in duplicate.
Superior Yield and Sensitivity
A: Comparison of efficiency. Amplify gDNA template with varying GC-content and length targets using each master mix’s recommended cycling conditions. 8 different targets were used. Ran in duplicate.
B: Comparison of yield. A gDNA template was amplified with varying GC-content and length targets using each master mix’s recommended cycling conditions. 8 different targets were used. The experiment was run in duplicate.
Long Amplification
A: Long Range Capabilities (gDNA). A range of 3.6 kb, 8.5 kb, 17.5 kb, and 24 kb gDNA templates were amplified with varying GC-content and length targets using each master mix’s recommended cycling conditions in a dye based qPCR. This experiment was run in duplicate.
B: Long Range Capabilities (λDNA). A range of 10 kb, 20 kb, 30 kb, and 40 kb λDNA templates were amplified with varying GC-content and length targets using each master mix’s recommended cycling conditions in a dye based qPCR. This experiment was run in duplicate.
Consistent GC Tolerance
Wide GC Tolerance Range. A gDNA template was amplified with varying GC-content and length targets using each master mix’s recommended cycling conditions. 8 different targets were used. The GC-content varied with 32%/900 base pairs (bp), 34%/800 bp, 38%/900 bp, 40%/1000 bp, 63%/300 bp, 64%/600 bp, 64%/800 bp, 70%/400 bp. This experiment was run in duplicate.
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