PPP Master Mix without MgCl2

Product description

PPP Master Mix without MgCl₂ is similar to PPP Master Mix (cat. No. P124 - P126) but does not contain MgCl₂, which is provided separately. This allows optimization of Mg²⁺ concentration, which is of key importance for proper PCR. The Mix contains Taq DNA polymerase, deoxyribonucleotides, reaction buffer components and additives.

Rapid samples preparation

• All components of the PPP Master Mix without MgCl₂ are 2x concentrated, which facilitates rapid preparation of the PCR samples. The samples are prepared by mixing an aliquot of PPP Master Mix with oligonucleotide primers, template DNA. MgCl₂ (included) and water (included).

Direct loading into the gel

• PPP Master Mix without MgCl₂ contains additives and a dye which allow direct loading of the samples into the gel, without necessity to add loading buffer.
• Dye present in the Mix migrates in the gel in front of the primers and therefore does not interfere with quantification of the PCR products. The dye and other additives have no effect on DNA amplification during PCR.

Optimized reaction buffer

• Reaction buffer in the Mix enhances specificity and efficiency of PCR; it contains a buffer, which after addition of proper amount of MgCl₂ is optimal for majority of PCRs.

• Stabilizers present in the Mix allow its storage for short periods at 4°C ± 3°C (days).

Technical data

Components and packaging

• 1 tube with 0.5 ml PPP Master Mix without MgCl₂ (for 40 reactions, 25 µl each)
• 1 tube with 0.5 ml 25 mM MgCl₂
• 1 tube with 1.5 ml PCR H₂O.

Composition      

• 2x concentrated PPP Master Mix without MgCl₂ contains: 150 mM Tris-HCl, pH 8.8 (at 25^oC), 40 mM (NH₄)₂SO₄, 0.02% Tween 20, 400 µM dATP, 400 µM dCTP, 400 µM dGTP, 400 µM dTTP, 100 U/ml Taq DNA polymerase, dye, stabilizers and additives.

Storage            

• For short terms (days) at 4°C ± 3°C.
• For long terms at -20 ± 5°C. Material can be repeatedly defrosted.

Purity and quality control:            

• Purity of Taq DNA polymerase is verified by SDS PAGE, only one band of 94 kDa is observed in Coomassie blue stained gel. Material is free of nucleases.
• Each batch of PPP Master Mix without MgCl₂ is tested for amplification of a single copy gene in genomic DNA.