"I used NEB Q5 for the same plasmid for PCR, same annealing temperature. Only the repliQa HiFi toughMix gave me the correct size products. It is very fast and works much better than NEB Q5 for difficult plasmids in PCR."
repliQa HiFi ToughMix
Features & Benefits
- 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
repliQa HiFi ToughMix is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
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.
Extreme Speed: Up to 3x faster resultsrepliQa HiFi ToughMix has fast extension times, ranging from 1-10 kb/sec depending on the fragment size, which significantly shortens time to results.
Extreme Speed: Up to 3x faster resultsComparison 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.
- Significantly faster results (up to 3x) with 1 kb/sec extension rate for fragments ≤1 kb
- No sacrifice in quality or yield of PCR results
Tough Tested: Tolerant to a wide range of PCR inhibitorsrepliQa HiFi ToughMix is able to tolerate a wide range of common PCR inhibitors, such as ethanol, hemoglobin and xylan, allowing for amplification of crude or difficult PCR sample types.
Tough Tested: Tolerant to a wide range of PCR inhibitorsStrong 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.
- Tolerant to a wide range of PCR inhibitors
- Amplify from crude samples
Superior Yield and SensitivityrepliQa HiFi ToughMix provides higher yield and sensitivity, highlighting the enzyme’s high efficiency. Coupled with extreme amplification speed, PCR products are able to be detected earlier and at lower levels.
Superior Yield and Sensitivity
Superior Yield and SensitivityA: 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.
- Consistently achieve higher yields across a range of inputs
- Greater sensitivity with low inputs
Long AmplificationrepliQa HiFi ToughMix has the ability to amplify long fragments of genomic DNA (+24 kb) and lambda DNA (+40 kb).
Long AmplificationA: 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.
- Amplify +24 kb gDNA and +40 kb lambda DNA
- Reduce time to result for longer fragments
Consistent GC TolerancerepliQa HiFi ToughMix is able to amplify varying levels of GC-content targets (32%-70% GC-rich), further enabling superior PCR performance with bacterial, plant and mammalian sample types.
Consistent GC Tolerance
Consistent GC ToleranceWide 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.
- Amplify varying levels of GC-content targets (32%-70%)
Contents2x reaction buffer containing optimized concentrations of MgCl2, dNTP’s and proprietarily formulated HiFi polymerase, hot start antibodies and ToughMix chemistry
Storage & HandlingStore kit components in a constant temperature freezer at -25°C to -15°C upon receipt.
For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.
Related ResourcesProduct FlyersProduct ManualsSafety Data Sheets (SDS)CofA (PSF)
"Worked great amplifying two gene target from plasmid DNA. It was the first time I have performed a two step reaction and it worked great. It was awesome running everything in 23 minutes too."