Taq DNA polymerases in which the phenylalanine is substituted by a tyrosine at position 667 (Taq F667Y) are members of a new class of DNA polymerases that incorporate chain-terminating dideoxyribonucleoside triphosphates (ddNTPs) much more efficiently than the wild-type Taq DNA polymerase. Improved incorporation of ddNTPs into DNA during cycle sequencing using AmpliTaq DNA polymerase, FS (Taq-F...

To achieve the production of a thermostable DNA polymerase free from bacterial DNA contamination, we developed eukaryote-made thermostable DNA (Taq) polymerase. The novel eukaryote-made thermostable DNA polymerase resolves the problem of contaminating bacterial DNA in conventional bacterially made thermostable DNA polymerase as a result of its manufacture and incomplete purification. Using euka...

DNA binding properties of the Type 1 DNA polymerases from Thermus aquaticus (Taq, Klentaq) and Escherichia coli (Klenow) have been examined as a function of [KCl] and [MgCl(2)]. Full-length Taq and its Klentaq "large fragment" behave similarly in all assays. The two different species of polymerases bind DNA with sub-micromolar affinities in very different salt concentration ranges. Consequently...

Thermostable DNA polymerase gene from Thermus aquaticus was cloned into constructed Taq from Thermus a Qaticus (pTTQ) plasmid using EcoRI and SalI sites with subsequent transformation in Escherichia coli strain (TOP10). The use of Isopropyl-β-Dthiogalactopyranosid (IPTG) as inducer of interested gene expression under control of the lac promoter was investigated. The optimization of enzyme induc...

background: taq dna polymerase is a very important enzyme for molecular biological studies such as dna amplification and dna sequencing by the pcr. it is a standard enzyme that is used in 90% of molecular biology labs today. the aim of this study was to produce taq dna polymerase enzyme in e. coli by a reliable, practical, simple and low cost method.materials and methods: in this study, the taq...

PCR co-amplification of two distinct HIV1 tat gene sequences lead to the formation of recombinant DNA molecules. The frequency of such recombinants, up to 5.4% of all amplified molecules, could be decreased 2.7 fold by a 6 fold increase in Taq DNA polymerase elongation time. Crossover sites mapped essentially to three discrete regions suggesting specific Taq DNA polymerase pause or termination ...

Few techniques are available to detect DNA lesions in cultured cells at the nucleotide level (8). One such method is primer extension of genomic DNA (5) that may be improved using linear amplification by repeated PCR cycles (1,11). This method has been used successfully to map the genomic sites of topoisomerase II (2,3,7) and topoisomerase I activity (10). DNA topoisomerases modulate DNA struct...

To investigate the influence of the pyrimidine 2-keto group on selection of nucleotides for incorporation into DNA by polymerases, we have prepared two C nucleoside triphosphates that are analogues of dCTP and dTTP, namely 2-amino-5-(2'-deoxy-beta-d-ribofuranosyl)pyridine-5'-triphosphate (d*CTP) and 5-(2'-deoxy- beta-d-ribofuranosyl)-3-methyl-2-pyridone-5'-triphosphate (d*TTP) respectively. Bot...