optimization of the condition for pcr-directed sequencing of microsatellites poly adenine (a) length polymorphisms is more difficult and sensitive compared with other common sequences. replication slippage may occur for polymerase enzyme during microsatellite amplification and direct sequencing of these pcr products will be challenging for heterozygote samples. so, the aim of this study is to i...

Modified PCR amplification and direct sequencing procedures for the double-stranded genomic DNA template are described. Advantages of the approach we describe are: background artifact bands previously observed using high-molecular-weight DNA as a template were eliminated by this protocol; no gel purification or subcloning of the PCR-amplified double-stranded fragment was required prior to direc...

The identification of the species of origin of meat and meat products is an important issue to prevent and detect frauds that might have economic, ethical and health implications. In this paper we evaluated the potential of the next generation semiconductor based sequencing technology (Ion Torrent Personal Genome Machine) for the identification of DNA from meat species (pig, horse, cattle, shee...

Direct DNA sequencing of amplified polymerase chain reaction (PCR) products offers several advantages over cloning of amplified DNA products. It is faster (1 day versus 3—5 days) and in DNA samples containing sequence polymorphisms both the normal and mutated sequence can be detected in the same sequencing reaction. The major problems encountered in direct sequencing of amplified DNA have been ...

A 72 kb region of BAC259.12D that encompasses the HAR1 locus was sequenced in its entirety using a DNA Sequencing Kit (PE Applied Biosystems) with an automated DNA sequencer (ABI PRISM 3100; PE Applied Biosystems). HAR1 complementary DNA was cloned from a cDNA library of L. japonicus shoots using DNA fragments of the first exon of the HAR1 gene obtained by polymerase chain reaction (PCR) from B...

AIM Rapid detection of H.pylori strains by PCR-Sequencing. METHODS 16S rDNA amplification by PCR from template genomic DNA, confirmation of amplicon size by agarose gel electrophoresis, sequencing of amplicons by automated sequencer, analysis of sequences by NCBI -BLAST software. RESULTS The PCR -Sequencing and analysis of the sequence data by BLAST resulted in detection of the strain to be...

Epigenetic changes of DNA, including methylation, have long been recognized as key indicators various diseases, aging, cancer, and neurological disorders. Biomarker discoveries based on distinct methylation patterns for both hypermethylation hypomethylation lead the way in discovery novel diagnosis treatment targets. Many different approaches are present to detect level whole genome (whole bisu...

Metagenomic analysis is the comprehensive study of DNA using clinical specimens organisms including bacteria, fungi, and viruses. In this study, we investigated efficacy metagenomic for diagnosing ocular infections, 11 keratitis cases, four iridocyclitis one endophthalmitis case. Corneal scraping, aqueous humor, vitreous were collected respectively. Ocular used bacterial fungal culture, PCR det...

In this Article the Authors incorrectly stated that they had developed a novel method termed bisulfite amplicon sequencing (BSAS). The method was reported by Masser et al. (reference 18 in the Article). Thus the following sentence (which appears verbatim in ref. 18): " By combining the benefits of bisulfite conversion, targeted amplification, tagmentation-based library construction , and NGS, w...