DNA polymerases contain active sites that are structurally superimposable and conserved in amino acid sequence. To probe the biochemical and structure-function relationship of DNA polymerases, a large library (200,000 members) of mutant Thermus aquaticus DNA polymerase I (Taq pol I) was created containing random substitutions within a portion of the dNTP binding site (Motif A; amino acids 605-6...

The RFLP/PCR approach (restriction fragment length polymorphism/polymerase chain reaction) to genotypic mutation analysis described here measures mutations in restriction recognition sequences. Wild-type DNA is restricted before the resistant, mutated sequences are amplified by PCR and cloned. We tested the capacity of this experimental design to isolate a few copies of a mutated sequence of th...

En 1978, une mission canadienne travaillant dans le complexe archeologique de Qal’eh-ye Yazdegerd a mene saison fouilles Gach Dawar ou les vestiges du chahar taq et des pieces adjacentes ont ete decouvertes. Les nouvelles 2007 permis reexaminer initialement reconnues lors canadiennes. La decouverte d’un certain nombre installations cultuelles l’interieur (une plateforme au centre laquelle il y ...

We investigated three probe design strategies used in quantitative polymerase chain reaction (PCR) for sensitivity in detection of the PCR amplicon. A plasmid with a 120-bp insert served as the DNA template. The probes were TaqMan, conventional molecular beacon (MB), and shared-stem molecular beacon (ATssMB and GCssMB). A shared-stem beacon probe combines the properties of a TaqMan probe and a ...

BACKGROUND AND AIMS Cholesteryl ester transfer protein (CETP) is an enzyme with a key role in lipoprotein metabolism. A common genetic polymorphism, the Taq 1B, influences CETP activity and HDL-cholesterol levels, with individual homozygotes for the B1 allele exhibiting higher enzyme activity and lower HDL-cholesterol levels than carriers of at least one B2 allele. Our aim was to analyze the in...

Three approaches were used to identify DNA polymorphisms in humans by examining the DNAs of a number of individuals within Huntington's Disease pedigrees as well as random individuals for restriction fragment length polymorphisms (RFLPs). The first approach used randomly chosen, cloned single-copy DNA segments from four types of human recombinant DNA libraries as hybridization probes to detect ...

The preparation of cDNA libraries usually involves multiple steps, such as isolation of poly A mRNA, synthesis of cDNA, cloning of cDNA into plasmid, and amplification of cloned plasmid. The last step, the amplification of insert DNA fragment after cloned in dephosphorylated plasmid vector, is done in either of two ways. The recombinant DNA can be extracted after amplification in host cells, or...

The conjunction of high resolution genetic maps based on (CA)n microsatellite markers (1) and fluorescent genotyping (2) has led to research programs which require the determination of hundreds of thousands of genotypes. However, the migration profile of a (CA)n microsatellite marker after PCR (polymerase chain reaction) is often complicated because of slippage of Taq polymerase during PCR, and...

As the sole renal Na,K-ATPase isozyme, the a, Na,K-ATPase accounts for all active transport of Na throughout the nephron. This role in renal Na reabsorption and the primacy of the kidney in hypertension pathogenesis make it a logical candidate gene for salt-sensitive genetic hypertension. An adenine (A)—>thymine (T) transversion, resulting in the substitution of glutamine with leucine and assoc...

The restriction enzyme TaqI digests 0.2% of the genomic DNA from the grasshopper Caledia captiva to a family of sequences 168 bp in length (length of consensus sequence). The sequence variation of this "Taq family" of repeat units was examined among four races from C. captiva to assay the pattern of evolution within this highly repeated DNA. The Taq-family repeats are located in C-banded hetero...