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repliQa HiFi ToughMix

Superior speed and inhibitor tolerance
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

 

*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.

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repliQa HiFi ToughMix
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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.

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What Customers Say


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.

Dr. Kara C.
UT Southwestern Medical Center

Details

Details

Contents

2x reaction buffer containing optimized concentrations of MgCl2, dNTP’s and proprietarily formulated HiFi polymerase, hot start antibodies and ToughMix chemistry

Customer Testimonials

Customer Testimonials

repliQa HiFi ToughMix

"The kit have a great power of amplification and extreme fast speed, I just need one hour to get result which I usually spent three hours before."

Simon S. | Professor, Tennessee State University
repliQa HiFi ToughMix

"This can amplify the tough template for SURE. I was struggled with the tough template that showed poor quality results with other many PCR enzymes. This repliQa saved my life and eventually gave me the clear and strong band by gel."

Lili C. | Post Doc, USC
repliQa HiFi ToughMix

"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."

Dr. Kara C. | UT Southwestern Medical Center
repliQa HiFi ToughMix

"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."

Dr. Brandon L. | University of Nebraska
repliQa HiFi ToughMix

"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."

Dr. Brandon L. | University of Nebraska

Details

Contents

2x reaction buffer containing optimized concentrations of MgCl2, dNTP’s and proprietarily formulated HiFi polymerase, hot start antibodies and ToughMix chemistry

Performance Data

Resources

Publications

A Non-Destructive High-Speed Procedure to Obtain DNA Barcodes from Soft-Bodied Insect Samples with a Focus on the Dipteran Section of Schizophora
Frederik Stein - 2022
Abstract
While the need for biodiversity research is growing, paradoxically, global taxonomical expertise is decreasing as a result of the neglected funding for young academics in taxonomy. Non-destructive approaches for DNA barcoding are necessary for a more efficient use of this dwindling expertise to fill gaps, and identify incorrect entries in sequence databases like BOLD or GenBank. They are efficient because morphological re-examination of species vouchers is still possible post-DNA barcoding. Non-destructive approaches for Diptera with a comprehensive species representation or the consideration of diagnostic fragile morphological characters are missing. Additionally, most non-destructive approaches combine a time intensive and non-destructive digestion step with common DNA extraction methods, such as commercial kits or CTAB DNA isolation. We circumvented those approaches and combined a modified non-destructive TE buffer high-speed DNA extraction, with a PCR inhibitor-resistant PCR reaction system, to a non-destructive DNA barcoding procedure for fresh and frozen samples of the Schizophora (Diptera). This method avoids morphological impairment and the application of harmful chemicals, is cost and time effective, restricts the need for laboratory equipment to a minimum, and prevents cross-contamination risk during DNA isolation. Moreover, the study indicates that the presented non-destructive DNA barcoding procedure is transferable to other soft-bodied insects. We suggest that PCR inhibitor-resistant master mixes enable the development of new—and the modification of existing—non-destructive approaches with the avoidance of further DNA template cleaning.
A Non-Destructive High-Speed Procedure to Obtain DNA Barcodes from Soft-Bodied Insect Samples with a Focus on the Dipteran Section of Schizophora
Frederick Stein - 2022
Abstract
While the need for biodiversity research is growing, paradoxically, global taxonomical expertise is decreasing as a result of the neglected funding for young academics in taxonomy. Non-destructive approaches for DNA barcoding are necessary for a more efficient use of this dwindling expertise to fill gaps, and identify incorrect entries in sequence databases like BOLD or GenBank. They are efficient because morphological re-examination of species vouchers is still possible post-DNA barcoding. Non-destructive approaches for Diptera with a comprehensive species representation or the consideration of diagnostic fragile morphological characters are missing. Additionally, most non-destructive approaches combine a time intensive and non-destructive digestion step with common DNA extraction methods, such as commercial kits or CTAB DNA isolation. We circumvented those approaches and combined a modified non-destructive TE buffer high-speed DNA extraction, with a PCR inhibitor-resistant PCR reaction system, to a non-destructive DNA barcoding procedure for fresh and frozen samples of the Schizophora (Diptera). This method avoids morphological impairment and the application of harmful chemicals, is cost and time effective, restricts the need for laboratory equipment to a minimum, and prevents cross-contamination risk during DNA isolation. Moreover, the study indicates that the presented non-destructive DNA barcoding procedure is transferable to other soft-bodied insects. We suggest that PCR inhibitor-resistant master mixes enable the development of new—and the modification of existing—non-destructive approaches with the avoidance of further DNA template cleaning.
A diverse group of underappreciated zygnematophytes deserves in-depth exploration
Anna Busch - 2022
Abstract
The conjugating green algae (Zygnematophyceae) are the closest relatives of land plants and hence are of great evolutionary interest. Besides the popular placoderm desmids and the filamentous species, there is an underappreciated diversity of unicellular zygnematophytes with a much “simpler” morphology and smooth cell walls – traditionally referred to as “saccoderm desmids”. These saccoderm desmids have a broad geographic distribution and are ecologically diverse. Many species inhabit terrestrial habitats such as dead wood, rock surfaces and glacial ice. Furthermore, several of the saccoderm genera have turned out to be highly polyphyletic and are typically poorly captured by environmental sequencing approaches. One of these genera is Mesotaenium Nägeli, with ~70 described species and infraspecific taxa united only by a relatively simple (plate- or ribbon-like) chloroplast structure. Here, we shed some light on these inconspicuous yet important members of the algal flora and present an updated rbcL gene phylogeny of the conjugating green algae, including several new lineages of Mesotaenium-like zygnematophytes. We depict the subtle morphological differences among these lineages and discuss our updated phylogeny in the light of ecology and cell biology. In addition, we review published knowledge on photoprotective strategies of zygnematophytes, the latest insights into their evolutionary innovations, and address some technical challenges in exploring this elusive group of microalgae. Some new observations of saccoderm desmids in undersampled habitats and of their microbial associates (e.g., parasites) point to interesting avenues for future research.
Opposite effects of stress on effortful motivation in high and low anxiety are mediated by CRHR1 in the VTA
Ioannis Zalachoras - 2022
Abstract
Individuals frequently differ in their behavioral and cognitive responses to stress. However, whether motivation is differently affected by acute stress in different individuals remains to be established. By exploiting natural variation in trait anxiety in outbred Wistar rats, we show that acute stress facilitates effort-related motivation in low anxious animals, while dampening effort in high anxious ones. This model allowed us to address the mechanisms underlying acute stress–induced differences in motivated behavior. We show that CRHR1 expression levels in dopamine neurons of the ventral tegmental area (VTA)—a neuronal type implicated in the regulation of motivation—depend on animals’ anxiety, and these differences in CRHR1 expression levels explain the divergent effects of stress on both effortful behavior and the functioning of mesolimbic DA neurons. These findings highlight CRHR1 in VTA DA neurons—whose levels vary with individuals’ anxiety—as a switching mechanism determining whether acute stress facilitates or dampens motivation.
Bacillus velezensis Identification and Recombinant Expression, Purification, and Characterization of Its Alpha-Amylase
Xiaodong Zhang - 2021
Abstract
Amylases account for about 30% of the global market of industrial enzymes, and the current amylases cannot fully meet industrial needs. This study aimed to identify a high α-amylase producing bacterium WangLB, to clone its α-amylase coding gene, and to characterize the α-amylase. Results showed that WangLB belonged to Bacillus velezensis whose α-amylase gene was 1980 bp coding 659 amino acids designated as BvAmylase. BvAmylase was a hydrophilic stable protein with a signal peptide and a theoretical pI of 5.49. The relative molecular weight of BvAmylase was 72.35 kDa, and was verified by SDS-PAGE. Its modeled structure displayed that it was a monomer composed of three domains. Its optimum temperature and pH were 70 °C and pH 6.0, respectively. It also showed high activity in a wide range of temperatures (40–75 °C) and a relatively narrow pH (5.0–7.0). It was a Ca2+-independent enzyme, whose α-amylase activity was increased by Co2+, Tween 20, and Triton X-100, and severely decreased by SDS. The Km and the Vmax of BvAmylase were 3.43 ± 0.53 and 434.19 ± 28.57 U/mg. In conclusion, the α-amylase producing bacterium WangLB was identified, and one of its α-amylases was characterized, which will be a candidate enzyme for industrial applications.

Customer Testimonials

repliQa HiFi ToughMix

"The kit have a great power of amplification and extreme fast speed, I just need one hour to get result which I usually spent three hours before."

Simon S. | Professor, Tennessee State University
repliQa HiFi ToughMix

"This can amplify the tough template for SURE. I was struggled with the tough template that showed poor quality results with other many PCR enzymes. This repliQa saved my life and eventually gave me the clear and strong band by gel."

Lili C. | Post Doc, USC
repliQa HiFi ToughMix

"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."

Dr. Kara C. | UT Southwestern Medical Center
repliQa HiFi ToughMix

"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."

Dr. Brandon L. | University of Nebraska
repliQa HiFi ToughMix

"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."

Dr. Brandon L. | University of Nebraska

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