2X qPCR-Mastermix Bio-Star for probes (LOW-ROX)

Features:

 - Enzyme with hot start capability increases reaction specificity and
   sensitivity, optimized

- DFS-Taq PLUS DNA polymerase activation requires not more than 5
  min heating

- High selectivity and reaction yield

- The mix is colored for easy pipetting

- Reduced preparation time

Applications:

- Real-time PCR with oligonucleotide probes

- Conventional PCR

- High-throughput PCR

- Genotyping

- For detection of bacterial DNA

Platforms:

Life Technologies (ABI) 7500, 7500 Fast, ViiA 7, QuantStudio 12K; Stratagene Mx4000, Mx3005P, and Mx3000P.

Description

Bio-Star qPCR-Mastermix (2×) LOW-ROX for probes is developed for quantitative real-time PCR with fluorescent probes. The Mastermix contains all components, except template and primers, for successful PCR.

- The mix is optimized for efficient and reproducible hot-start real-time
   PCR of genomic, plasmid and viral DNA samples.

- The solution contains substances that increase half-life and
  processivity of DFS-Taq PLUS DNA polymerase blocked by MAB amd
  by enhancing its stability during PCR.

- It includes components that influence primer annealing temperature
  and characteristics of template melting thus enabling to increase the
  specificity of PCR and use templates with complicated structure.

- DFS-Taq Plus DNA polymerase is inactive at room temperature
  because of monoclonal antibodies.

Components and Mixture

2X Bio-Star qPCR-Mastermix LOW-ROX for probes contains:

- 100 mM Tris-НCl (рН 8.5 at 25 °С), 100 mM KCl, 0.4 mM each of
  dNTP, 10 mM MgCl2, 0.1 U/µl DFS-Taq DNA Polymerase, 0.025%
  Tween 20,  60 nM ROX fluorescent dye, stabilizers and enhancers,

- Tube of MgCl2 100 mM

- Water Mol.Bio Grade 2 x 1,25 ml

Storage and transportation: at -20 °С; not more than 50 thawing-freezing cycles. shipping with blue ice or at room temperature

Storage terms: up to 24 mounts

Exodeoxyribonuclease activity: DNA was stable after incubation of 1 µg fragment of phage lambda DNA in the presence of 25 µl of Bio-Star qPCR Mastermix (2×) in 50 µl reaction solution at 37 °C and 70 °C for 4 h.

Test for bacterial DNA contamination

Test for Hot -Start ( we test activity with or without MAB at various temperatures

Exodeoxyribonuclease activity

DNA was stable after incubation of 1 μg fragment of phage lambda DNA in the presence of 25 μl of BioMaster HS-qPCR (2×) in 50 μl reaction solution at 37 °C and 70 °C for 4 h.

Ribonuclease activity

Absence of ribonuclease activity was confirmed after incubation of 1 μg of 5’-[P32]-labeled RNA fragment in the presence of 25 μl of BioMaster HS-qPCR (2×) in 50 μl reaction solution at 37 °C for 4 h.

Stability:

One month at room temperature does not reduce PCR efficiency

1. Defrost the reaction mixture and stir thoroughly.

2. Add the following components into the thin-wall PCR tubes considering the final volume of a reaction mixture equal to 50 μl: