Screening for mutations in human disease-causing genes in a molecular diagnostic environment demands simplicity with a view to allowing high throughput approaches. In order to advance these requirements, we have developed and applied a primer design program, termed BatchPD, to achieve the PCR amplification of coding exons of all known human Refseq genes. Primer design, in silico PCR checks and ...

A simple method, primer specific and mispair extension analysis (PSMEA) with pfu DNA polymerase was developed for genotyping. PSMEA is based on the unique properties of 3'-->5' exonuclease proofreading activity. In the presence of an incomplete set of dNTPs, pfu was found to be extremely discriminative in nucleotide incorporation and proofreading at the initiation step of DNA synthesis, complet...

To aid in the characterization of the relationship of structure and function for human immunodeficiency virus type-1 reverse transcriptase (HIV-1 RT), this investigation utilized DNAs containing benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE)-modified primers and templates as a probe of the architecture of this complex. BPDE lesions that differed in their stereochemistry around the C10 posit...

Salmonella enterica is a significant cause of gastroenteritis worldwide, with serovars Typhimurium and Heidelberg being particularly prevalent, which have broad host ranges infecting poultry, dairy animals, and humans. Traditional methods used for the detection of Salmonella from contaminated food products are time-consuming and labor-intensive. The aim of this study was to develop a sensitive ...

Aptamers are oligonucleotides that can bind to specific target molecules. Most aptamers are generated using random libraries in the standard systematic evolution of ligands by exponential enrichment (SELEX). Each random library contains oligonucleotides with a randomized central region and two fixed primer regions at both ends. The fixed primer regions are necessary for amplifying target-bound ...

Recently, we developed a simple isothermal nucleic acid amplification reaction, primer generation-rolling circle amplification (PG-RCA), to detect specific DNA sequences with great sensitivity and large dynamic range. In this paper, we combined PG-RCA with a three-way junction (3WJ) formation, and detected specific RNA molecules with high sensitivity and specificity in a one-step and isothermal...

MOTIVATION DNA methylation plays important roles in biological processes and human diseases, especially cancers. High-throughput bisulfite genomic sequencing based on new generation of sequencers, such as the 454-sequencing system provides an efficient method for analyzing DNA methylation patterns. The successful implementation of this approach depends on the use of primer design software capab...

A primer set of oligonucleotides (S18 and S19) from the ompC gene of Salmonella has been evaluated for specific detection of Salmonella by polymerase chain reaction (PCR). This primer set successfully amplified 40 Salmonella serovars (60 isolates), but not 24 non-Salmonella bacteria (42 isolates) that have been tested so far. The uniqueness of these primer sequences was also confirmed. The sens...

Characterization of synthetic oligonucleotides and quantification of primer extension mediated by a human translesion synthesis polymerase η (Pol η) over drug-induced DNA lesions in the presence on modified nucleotide analogs is described. Extent of primer extension for each reaction was monitored by denaturing gel electrophoresis. The data was obtained to assess the performance of the fluoresc...

Ayu (Plecoglossus altivelis), one kinds of valuable cultured fish species, show almost no morphological difference between male and female until sexual maturity. Here, we report the identification of sex-linked markers for the ayu, based on Amplified Fragment Length Polymorphism (AFLP) generated from cultured fish (15 males and 15 females) by using 63 different primer combinations. Genomic frag...