PCR-Mastermix is a premixed ready-to-use solution containing all reagents required for PCR (except template, primers and water).
Reagent Composition PCR-Mastermix:
- 5 x Reaction Buffer B (400 mM Tris-HCl pH 9,4 – 9,5 at 25°C, 100 mM (NH4)2SO4, 0,1%
- 12,5 mM MgCl2, (= 2,5 mM in 1x solution)
- 1 mM dNTPs of each : 1 x PCR solution – dATP, dGTP, dCCT und dTTP (= 200 µM/dNTP in
Source: Purified from an E.coli strain carrying Taq-DNA-Polymerase overproducing plasmid. The original enzyme has been isolated from Thermus aquaticus.
Associated activities: Taq-DNA-Polymerase is a highly processive 5’ --> 3’-DNA-polymerase with 5’ --> 3’-exonuclease activity. 3’ --> 5’-exonuclease activity lacks completely. Additionally, the enzyme adds nucleotides (mostly adenosines) to the 3´-ends of the DNA, so that TA-cloning is possible without further modifications.
Application & Quality control:
Primer extension reaction: the enzyme is free of nicking and primer extension activities as well as of exonucleases and unspecific endonucleases. SDS/PAGE: 95 kD-band, purity >98%. Activity and stability tested via PCR. The error rate per nucleotide per cycle is ~8,3x10 -5; the accuracy ~ 1,2x10 4. Estimated half life at 95ºC: 90 min.
Volume: 1000 µl or 5000 µl