Pfu高保真热启动聚合酶
A high-fidelity DNA polymerase (higher than 130x Taq fidelity), High-Specificity Pfu HS Mix is highly suited to DNA target enrichment, NGS library amplification, and cloning applications. High-Specificity Pfu HS Mix provides high yield and sensitivity, as well as superior amplification of GC- and AT-rich targets. The hot-start aptamer binds reversibly to the high-fidelity DNA polymerase which inhibits its activity at ambient temperatures and improves the specificity of the amplification.
High-Specificity Pfu HS Mix, MDX006
Available in 5 mL (200 Units) or 100 mL (4,000 Units) aliquots
High-Specificity Pfu HS Mix is a combination of the latest advances in buffer chemistry and PCR enhancers and stabilizers, together with an aptamer-mediated hot-start polymerase, dNTPs, and MgCl2. The advanced buffer chemistry has been developed for fast, highly reproducible, multiplex PCR and is designed for superior sensitivity and specificity, making High-Specificity Pfu HS Mix perfect for DNA target enrichment and NGS library amplification.
产品资料
High-Specificity Pfu HS CatalogHigh-Specificity Pfu HS Catalog
分子诊断的原料试剂解决方案分子诊断的原料试剂解决方案
FAQs: High-Specificity Pfu HS Mix
在自然界中，DNA聚合酶在DNA链延长过程中纠错核苷酸整合的能力对于个体生存至关重要。这被称为“校对活动”，发生在3′到5′方向。聚合酶的这种活性还能去除3'端（A-端）的未配对核苷酸，产生钝端。
高特异性Pfu HS Mix保真度的测定采用使用条形码策略和NGS来描述DNA聚合酶的平均错误率、错误热点和病变绕过的保真度。 ( Mapping DNA polymerase errors by single-molecule sequencing. Lee F. D., et al. Nucleic Acids Research, 44 (13) e118 2016 (doi: 10.1093/nar/gkw436))
高特异性Pfu HS Mix可用于含有PCR抑制物的样本中。增加镁离子浓度可以避免反应中过多的干扰，例如，在10%全血样本中，可以将镁离子浓度增加到4mM。
高特异性Pfu HS Mix已经验证了可用于长达5 kb的模板。
DNA target enrichment is a pre-sequencing DNA preparation step where DNA sequences are either directly amplified (multiplex PCR-based) or captured (hybrid capture-based). These enriched DNA fragments can then be sequenced using DNA sequencers.
Aptamers are engineered oligonucleotides that bind to the active site of the high-fidelity DNA polymerase but are denatured and released as the temperature is increased during the PCR amplification cycle. This aptamer-mediated inhibition/activation process is however fully reversible, and so at the end of thermal cycling, when the temperature of the reaction is decreased, the aptamer refolds and rebinds to high-fidelity DNA polymerase, inhibiting any further activity in the sample. This has proven to be important in workflows where undesired polymerase activity after reaction completion can disturb baseline readings, such as NGS following amplification of a library.
不会，适配子的末端被阻断以防止它们形成序列。
高特异性Pfu HS Mix应保存在-20°C保留最佳活性。 不建议将高特异性Pfu HS Mix储存在-80°C，因为活性部位可能会形成冰晶，从而额响或破坏酶的活性
Specifications:
Description: High-fidelity PCR master mix containing a reversible, temperature-dependent hot start Taq DNA polymerase and buffer
Concentration: 2 x
Appearance: Clear, colorless solution
Hot Start. Aptamer mediated
Application: PCR, NGS target enrichment, NGS library preparation
Sample type: cDNA, DNA
Presentation: 1 vial
Storage: -20 °C
Mix stability: See outer label
Consistency: ± 0.5 Ct variance between test and reference sample
DNA Contamination: None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets.
DNase Contamination: No detectable degradation
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