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
Comparison of yield. A gDNA template was amplified with varying GC-content and length targets using each manufacturer’s recommended cycling conditions. The experiment was run using 8 different targets, in duplicate.
Long Amplification and Consistent GC Tolerance
A Long Range Capabilities. A range from 0.25 to 24 kb of gDNA templates were amplified with varying GC-content and lengths using recommended cycling
conditions. B/C Wide GC-content tolerance range. gDNA templates were amplified with varying GC-content and lengths using recommended cycling conditions.
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Customer Product Reviews
| 5 star | 65% | |
| 4 star | 27% | |
| 3 star | 4% | |
| 2 star | 2% | |
| 1 star | 0% |
repliQa HiFi ToughMix
The product is easy to use, the recommended PCR mixture and thermocycler protocol were helpful and effective.
I got a chance to test the RepliQa polymerase on some gDNA from our organism, which is GC rich and tends to give us problems amplifying using Q5.
I tried some reactions that didn’t work with Q5 (two 5kb reactions) and a big amplicon product of 11kb.
All my reactions worked great! There were some off target bands that were smaller than my expected band and just as bright as the expected size band, but we also see this with Q5 when it works. I gel extracted my products (two 5kb and one 11kb product) and sequenced these products and all aligned perfectly with no SNPs.
I also noticed that even if my primers were lower than 64ºC, the two step cycling worked while the 3 step didn’t.
This enzyme is wonderful because it gives us a second option when Q5 doesn’t work, but I think the price prevents us from completely replacing Q5. We plan to have repliQa in our lab as a plan B DNA polymerase.
This is by far the most robust and fastest polymerase I have used. I like the master mix format to ease of sample set up. reliQa HiFi ToughMix when tested head-to-head with other high-fidelity polymerase, performed better in higher and faster amplification. It was also able to amplify some of our tougher GC-rich constructs without any problem.