Direct Blood PCR Kit enables to perform PCR reactions directly from whole blood with no prior DNA extraction or purification.
The Direct Blood PCR Kit allows to use whole blood stored at +4°C or -20°C as well as dried blood spots on cards such as commercially available Whatman FTA® cards.
Blood can be preserved with all most often used anticoagulants: EDTA, citrate or heparin.
It is possible to use whole blood in a wide range of concentrations: from 1 % to 20 %. The recommended starting point is 5 % (MgCl2 concentration contained in the Blood PCR buffer is optimized for 5 % blood).
The Direct Blood PCR Kit employs a genetically engineered thermophilic DNA polymerase with high tolerance to blood inhibitors.
Blood DNA Polymerase catalyzes the polymerization of nucleotides into duplex DNA in the 5’۳′ direction.The enzyme exhibits the 3’۵′ proofreading activity, resulting in over 10-fold higher PCR fidelity than possible with Taq DNA Polymerases.
The enzyme generates blunt ends.
Enhanced polymerase processivity allows to use shorter extension times.
Due to the genetic modification of the polymerase, the optimal reaction conditions (especially annealing temperatures) differ from standard PCR protocols.
The Direct Blood PCR Kit allows to obtain a wide range of product size (over 4 kb).
Blood DNA Polymerase Storage Buffer: ۲۰ mM Tris-HCl (pH 8.0 at 22°C), 100 mM KCl, 0.5% Tween™۲۰, ۰٫۵% Igepal CA-630, 0.1 mM EDTA, 1 mM dithiothreitol, 50% glycerol and stabilizers.
۵ x Blood PCR Buffer: The buffer contains 15 mM MgCl2.
Quality Control: All preparations are assayed for contaminating 3′-exonuclease, and nonspecific single- and double-stranded DNase activities. Typical preparations are greater than 95% pure, as judged by SDS polyacrylamide gel electrophoresis.