Neq DNA polymerase is the first archaeal family B DNA polymerase reported to lack uracil recognition function and successfully utilize deaminated bases. We have focused on two amino acid residues (Y515, A523) in the fingers subdomain of Neq DNA polymerase, which were predicted to be located in the middle of the fingers subdomain, based on amino acid sequence alignment of the Neq DNA polymerase ...

a highly efficient cloning vector was constructed for cloning pcr products by inserting an 80 bp dna fragment into pgem-5zf (+) vector. the xcm i digestion of this vector gave rise to a 3’ overhanging deoxythymidine offering the possibility of cloning pcr products with 3' adenosine overhang created by taq dna polymerase. furthermore, two ecor i sites were added to the construct for identificati...

A simple and accurate genomic primer extension method has been developed to detect ultraviolet footprinting patterns of regulatory protein-DNA interactions in mammalian genomic DNA. The technique can also detect footprinting or sequencing patterns introduced into genomic DNA by other methods. Purified genomic DNA, containing either damaged bases or strand breaks introduced by footprinting or se...

Point mutations in somatic cells play a role in the etiology of several classes of human pathologies. Experimental procedures are required that allow the detection and quantitation of such mutations in disease-related genes in tissue biopsy samples without the need for the selection of mutated cells. We describe the genotypic analysis of single base pair mutations in the Taq I endonuclease reco...

Random amplified polymorphic DNA (RAPD) technique is a simple and reliable method to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of assay. In this study, we analyzed the effect of changing the concentration of primer, magnesium chloride, template DNA and Taq DNA polymerase with the objective of determining t...

Materials Lambda exonuclease ( exo), Exonuclease I (exo I), Lambda Exonuclease Buffer and ThermoPolReaction Buffer were all purchased from New England Biolabs (MA, USA). Taq DNA polymerase was purchased from Tiangen Biotech Co. (Beijing, China). DNA strands were synthesized and purified by HPLC (Sangon Co., China). The sequences of all the probes and targets that have been studied in this work...