Direct Blood PCR Kit enables to perform PCR reactions directly from whole blood with no prior DNA extraction or purification.
Description:
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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.
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Blood can be preserved with all most often used anticoagulants: EDTA, citrate or heparin.
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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).
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The Direct Blood PCR Kit employs a genetically engineered thermophilic DNA polymerase with high tolerance to blood inhibitors.
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Blood DNA Polymerase catalyzes the polymerization of nucleotides into duplex DNA in the 5'3' direction.The enzyme exhibits the 3'5' proofreading activity, resulting in over 10-fold higher PCR fidelity than possible with Taq DNA Polymerases.
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The enzyme generates blunt ends.
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Enhanced polymerase processivity allows to use shorter extension times.
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Due to the genetic modification of the polymerase, the optimal reaction conditions (especially annealing temperatures) differ from standard PCR protocols.
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The Direct Blood PCR Kit allows to obtain a wide range of product size (over 4 kb).
Blood DNA Polymerase Storage Buffer: 20 mM Tris-HCl (pH 8.0 at 22°C), 100 mM KCl, 0.5% Tween™20, 0.5% Igepal CA-630, 0.1 mM EDTA, 1 mM dithiothreitol, 50% glycerol and stabilizers.
5 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.