BioMaster UDG HS-qPCR Lo-ROX (2×)

Features
2× reaction mix containing Hot-Start Taq DNA polymerase, dUTP and N-uracil-DNA glycosylase for real-time PCR with fluorescent probes; contains ROX at low concentration for studies involving use of specific equipment.
Application
  • Hot-start real-time PCR with fluorescently labeled probes; the mix is recommended for amplifiers with a channel for ROX registration
  • Conventional PCR
  • High-throughput PCR
  • Multiplex PCR
  • Genotyping

Basic description

BioMaster UDG HS-qPCR Lo-ROX (2×) contains 2× BioMaster UDG HS-qPCR Lo-ROX reaction mix and sterile water. The mix is developed for quantitative real-time PCR with fluorescently labeled probes on PCR platforms that utilize ROX as a reference dye. BioMaster UDG HS-qPCR Lo-ROX (2×) includes all components (except for DNA template, primers and probe) necessary for PCR: highly processive recombinant HS-Taq DNA polymerase, N-uracil-DNA glycosylase, dNTP mix, PCR buffer, Mg2+ solution, and ROX dye.

BioMaster UDG HS-qPCR Lo-ROX (2×) is optimized for efficient and reproducible hot-start real-time PCR of genomic, plasmid and viral DNA samples. The mix is supplemented with additives increasing half-life and processivity of HS-Taq DNA polymerase by enhancing its stability during PCR. BioMaster UDG HS-qPCR Lo-ROX (2×) mix does not contain substances that influence primer annealing temperature and characteristics of template melting.

N-uracil-DNA glycosylase (UDG) and dUTP (in proportion with dTTP) provide protection from amplicon transfer between reaction solutions (cross-contamination). DNA polymerase included in the mix is inactive at room temperature, its activation requires preheating at 95 °C for 5 min.

The master mix is ideally suitable for PCR platforms that use ROX passive dye as a reference guide, such as Life Technologies (ABI) 7500, 7500 Fast, ViiA 7, QuantStudio 12K; Stratagene Mx4000, Mx3005P, and Mx3000P. Use of the kit saves time and minimizes contamination risk due to reduced number of pipetting steps.

Areas of use

  • Classic PCR with "hot" start
  • Genotyping
  • PCR in real time with "hot" start, with probes and intercalating dyes
  • PCR multiplex

Reaction mixture properties

  • The mix is inactive at room temperature due to the Hot-Start technology and is activated after incubation at 95 °C for 5 min
  • Allows normalization using ROX dye as a reference
  • Prevents re-amplification of PCR carryover products
  • The mix is optimized for the specific performance of HS-Taq DNA polymerase, long-term storage (the storage of BioMaster UDG HS-qPCR Lo-ROX (2×) at room temperature for 7 days does not affect PCR efficiency), multiple thawing-freezing cycles

Benefits of using

  • Enzyme with hot-start capability increases reaction specificity
  • Activation of HS-Taq DNA polymerase requires not more than 5 min heating
  • High selectivity and reaction yield
  • Reduced preparation time
  • Protection from cross-contamination
  • Allows data normalization
  • Standardized conditions of the same-type reactions (reduced pipetting error during mixing PCR components from experiment to experiment)

Storage

1 year (under proper storage and transportation conditions).

Transportation

at -20 °С; not more than 50 thawing-freezing cycles.

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