qPCR 2x Blue Master Mix

Product description

This product is an alternative to qPCR 2x Master Mix with different buffer, containing ammonium sulfate. This buffer under certain conditions enhances amplification, due to optimized concentration of Mg²⁺. The mix is used mostly for qPCR with DNA specific probes such as TaqMan, Molecular beacons, FRET and others.

Rapid preparation (2x concentrated)

• All components of the qPCR 2x Blue Master Mix are 2x concentrated (optimized reaction buffer, nucleotides, Taq DNA polymerase and anti-Taq monoclonal antibody). This facilitates rapid preparation of the PCRs. The samples are prepared simply by mixing an aliquot of the Mix with oligonucleotide primers, template DNA, H₂O (included) and selected DNA probes.

Hot start

• The product contains monoclonal antibody anti-Taq, which binds to Taq DNA polymerase and thus inactivates its enzymatic activity.  After the first denaturation cycle, the antibody is irreversibly inactivated and Taq DNA polymerase regains enzymatic activity. This decreases formation of nonspecific DNA amplicons.

Sensitive detection

• This product is optimized for sensitive detection of DNA fragments amplified during qPCR from genomic DNA or cDNA obtained by reverse transcription.

Rapid setup

• qPCR 2x Blue Master Mix is especially useful for routine analyses of large numbers of DNA samples. To 0.5 ml of the Master Mix in original tube, primers (e.g. 40 µl forward and 40 µl reverse), PCR H₂O and fluorescent probes are added and mixed; the "armed Mix” can be stored at -20 ± 5^oC. Immediately before PCR, the Mix is thawed and each 24 µl aliquot is mixed with 1 µl of the tested DNA template.

Technical data

Components and packaging

• 1 tube with 0.5 ml qPCR 2x Blue Master Mix (for 40 reactions, 25 µl each).
• 1 tube with 1.5 ml PCR H₂O.

Storage          

• At temperature -20^oC ± 5°C. Material can be repeatedly defrosted.

Composition  

• The Mix is 2x concentrated: 150 mM Tris-HCl, pH 8. 8 (at 25^oC), 40 mM (NH₄)₂SO₄, 0.02% Tween 20, 5 mM MgCl₂, 400 µM dATP, 400 µM dCTP, 400 µM dGTP, 400 µM dTTP, Taq DNA polymerase (50 U/ml), monoclonal antibody anti-Taq, stabilizers and additives.

Purity and quality control   

• Each batch of qPCR 2x Blue Master Mix is tested for amplification of a single copy gene in genomic DNA.