AccuStart Taq DNA Polymerase HiFi

Features & Benefits

  • Superior assay sensitivity and specificity with AccuStart enzyme technology – maximum-yielding Taq DNA polymerase mutant with stringent, multi-epitope antibody hot start
  • Optimized high-fidelity PCR buffer with 3’-exonuclease (proofreading) activity supports robust amplification of large PCR product ≥20kb in length

 

AccuStart Taq DNA Polymerase HiFi is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description

AccuStart Taq DNA Polymerase HiFi combines high-yielding mutant Taq DNA polymerase with 3’-exonuclease (proof-reading) polymerase and ultrapure, monoclonal antibody hot-start activation control. This reagent provides highly-sensitive and precise target amplification with convenient assembly at ambient temperature. Robust and reliable amplification of large, complex DNA targets up to 20kb in length.

Details

  • Contents
    • AccuStart Taq DNA polymerase HiFi 5 units/µL in 50% glycerol, 20 mM Tris-HCl, 40 mM NaCl, 0.1 mM EDTA, and stabilizers.
    • HiFi PCR Buffer (10X) 600 mM Tris-SO4 (pH 8.9), 180 mM (NH4)2SO4
    • 50 mM magnesium sulfate 50 mM MgSO4
  • Storage & Handling
    Store 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 Resources
    Product Manuals
    Safety Data Sheets (SDS)
    CofA (PSF)
    Publications
    KRAS mutations in blood circulating cell-free DNA: a pancreatic cancer case-control study
    Florence Le Calvez-Kelm, Impact Journals - 2016
    Abstract
    The utility of KRAS mutations in plasma circulating cell-free DNA (cfDNA) samples as non-invasive biomarkers for the detection of pancreatic cancer has never been evaluated in a large case-control series. We applied a KRAS amplicon-based deep sequencing strategy combined with analytical pipeline specifically designed for the detection of low-abundance mutations to screen plasma samples of 437 pancreatic cancer cases, 141 chronic pancreatitis subjects, and 394 healthy controls. We detected mutations in 21.1% (N=92) of cases, of whom 82 (89.1%) carried at least one mutation at hotspot codons 12, 13 or 61, with mutant allelic fractions from 0.08% to 79%. Advanced stages were associated with an increased proportion of detection, with KRAS cfDNA mutations detected in 10.3%, 17,5% and 33.3% of cases with local, regional and systemic stages, respectively. We also detected KRAS cfDNA mutations in 3.7% (N=14) of healthy controls and in 4.3% (N=6) of subjects with chronic pancreatitis, but at significantly lower allelic fractions than in cases. Combining cfDNA KRAS mutations and CA19-9 plasma levels on a limited set of case-control samples did not improve the overall performance of the biomarkers as compared to CA19-9 alone. Whether the limited sensitivity and specificity observed in our series of KRAS mutations in plasma cfDNA as biomarkers for pancreatic cancer detection are attributable to methodological limitations or to the biology of cfDNA should be further assessed in large case-control series.
    Sojun Hoshimoto, Journal of Thoracic Oncology: Official Publication of the International Association for the Study of Lung Cancer - 2015
    Abstract
    INTRODUCTION: Esophageal squamous cell carcinoma (ESCC) is a cancer of variable outcomes with limited effective treatments resulting in poor overall survival (OS). Epigenetic alterations contributing to this deadly cancer type that can be used as novel therapeutic or diagnostic targets are still poorly understood. METHODS: We explored genome-wide DNA methylation data from The Cancer Genome Atlas project and identified a panel of tumor-related genes hypermethylated in ESCC. The methylation statuses of RASSF1- RARB- CDKN2A (p16INK4a- p14ARF)- APC- and RUNX3 genes and long interspersed nucleotide element-1 (LINE-1) were validated in a large cohort (n = 140) of clinically well-annotated ESCC specimens and esophageal normal mucosa (n = 28) using a quantitative methylation-specific polymerase chain reaction. RESULTS: Hypermethylation of RARB- p16INK4a- RASSF1- APC- RUNX3- and p14ARF were observed in 55\%- 24\%- 20\%- 19\%- 14\%- and 8\% of specimens- respectively. Hypermethylation of APC was significantly associated with tumor depth (p = 0.02) and American Joint Committee on Cancer stage (p = 0.03). Global DNA methylation level- assessed by LINE-1- was significantly lower in ESCC than in normal mucosa (p {\textless} 0.0001)- and lower in greater than or equal to T2 (n = 69) than T1 tumors (n = 45, p = 0.03). There was a significant inverse correlation between LINE-1 and RARB methylation (p = 0.008). Importantly- hypermethylation of RASSF1 and APC genes was significantly associated with overall survival (OS, p = 0.006 and p = 0.007- respectively). In addition- patients with tumors containing a higher number of methylated genes (greater than two genes) presented worse OS (p = 0.003). CONCLUSIONS: This study demonstrates that epigenetic alterations of a panel of tumor-related genes and the noncoding region LINE-1 can be used as prognostic indicators and help in clinical management of ESCC patients.
    Primers for 52 polymorphic regions in the Quercus rubra chloroplast- 47 of which amplify across 11 tracheophyte clades
    Daniel Borkowski, Tree Genetics & Genomes - 2014
    Abstract
    Postglacial migration studies in Quercus rubra L. (northern red oak) are hampered by low levels of population differentiation in the widely used universal chloroplast primers. We sequenced the large single copy (LSC) regions of the Q. rubra and Quercus ellipsoidalis chloroplasts to enable us to query additional regions for future studies on migration and speciation. Using 454 sequencing of long-range PCR amplicons and Sanger sequencing for gap closure- we report 65 coding sequences from Q. rubra and 59 from Q. ellipsoidalis. Comparison of our de novo assembly of the LSC region sequence for Q. rubra to Q. rubra chloroplast sequence (NCBI Reference Sequence: NC\_020152.1) from a different tree revealed 106 polymorphisms- all within intergenic regions- that can serve as tools for postglacial migration studies and taxonomic studies within the Lobatae. Sequence alignment for the 59 complete coding regions in common for theQ. rubrachloroplast reference sequence- our Q. rubra sequence and our Q. ellipsoidalis sequence revealed no sequence polymorphisms and no indels. We also report the 52 primer pairs we used for gap closure- including 53 new primer pairs not previously reported. We tested these 52 primer pairs against 11 species representing the Tracheophyta and detected 47 that produced amplicons in all 11 species. The new universal primers we have identified provide additional tools for resolving the taxonomic relationships among the congeneric taxa of forest trees in the temperate and subtropical forests of the Northern Hemisphere.
    Comparative Genomics of cpn60-Defined Enterococcus hirae Ecotypes and Relationship of Gene Content Differences to Competitive Fitness
    Isha Katyal, Microbial Ecology - 2015
    Abstract
    Natural microbial communities undergo selection-driven succession with changes in environmental conditions and available nutrients. In a previous study of the pig faecal Enterococcus community, we demonstrated that cpn60 universal target (UT) sequences could resolve phenotypically and genotypically distinct ecotypes of Enterococcus spp. that emerged over time in the faecal microbiome of growing pigs. In this study, we characterized genomic diversity in the identified Enterococcus hirae ecotypes in order to define further the nature and degree of genome content differences between taxa resolved by cpn60 UT sequences. Genome sequences for six representative isolates (two from each of three ecotypes) were compared. Differences in phosphotransferase systems and amino acid metabolism pathways for glutamine, proline and selenocysteine were observed. Differences in the lac family phosphotransferase system corresponded to lactose utilization phenotypes of the isolates. Competitive fitness of the E. hirae ecotypes was evaluated by in vitro growth competition assays in pig faecal extract medium. Isolates from E. hirae-1 and E. hirae-2 ecotypes were able to out-compete isolates from the E. hirae-3 ecotype, consistent with the relatively low abundance of E. hirae-3 relative to E. hirae-1 and E. hirae-2 previously observed in the pig faecal microbiome, and with observed differences between the ecotypes in gene content related to biosynthetic capacity. Results of this study provide a genomic basis for the definition of ecotypes within E. hirae and confirm the utility of the cpn60 UT sequence for high-resolution profiling of complex microbial communities.
    A Universal Approach to Prepare Reagents for DNA-Assisted Protein Analysis
    Junhong Yan, PLoS ONE - 2014
    Abstract
    The quality of DNA-labeled affinity probes is critical in DNA-assisted protein analyses- such as proximity ligation and extension assays- immuno-PCR- and immuno-rolling circle amplification reactions. Efficient- high-performance methods are therefore required for isolation of pure conjugates from reactions where DNA strands have been coupled to antibodies or recombinant affinity reagents. Here we describe a universal- scalable approach for preparing high-quality oligonucleotide-protein conjugates by sequentially removing any unconjugated affinity reagents and remaining free oligonucleotides from conjugation reactions. We applied the approach to generate high-quality probes using either antibodies or recombinant affinity reagents. The purified high-grade probes were used in proximity ligation assays in solution and in situ- demonstrating both augmented assay sensitivity and improved signal-to-noise ratios.
    Click here to see all Publications

Ordering Information