Bio-Star Mastermix (2X) High-ROX

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
- Reduced preparation time


Applications:
- Real-time PCR with oligonucleotide probes
- Conventional PCR
- High-throughput PCR
- Genotyping
- For detection of bacterial DNA


Platforms:
(ABI) 7000, 7300, 7700, 7900, 7900HT and StepOnePlus

Description
Bio-Star PCR-Mastermix (2×) HIGH-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 PCR-Master mix HIGH-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, 900 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.
 
Stability tests / Quality control / Comparison

• 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