Polymerase Fidelity Testing at NEB is based on error accumulation in the lacZ gene during PCR. A technique developed by Wayne Barnes.
First, the lacZ gene is amplified by PCR with the polymerase of interest, leading to the accumulation of errors at the polymerase's intrinsic rate of error incorporation. The amplified DNA is digested by the restriction enzymes NcoI and HindiIII, and inserted into a plasmid while restoring two antibiotic resistance genes.
The plasmids are then transformed into competent E. coli. The E. coli are either grown on selective media containing X-gal, so that blue and white colonies can be counted, or grown on selective media, isolated, then sequenced.
Help us celebrate our 50th anniversary! We have hidden 1,000 golden butterflies and are waiting for you to find them. They can be anywhere that you find NEB! Beginning April 15th, be sure to visit our website and tables at tradeshows and events you are attending. Visit our social media channels frequently for tips on where we have hidden the butterflies – and once you find one, either click or scan the code to be eligible for a 50th anniversary prize pack, as well as a grand prize trip to NEB headquarters in Ipswich, MA!.
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