The determination of unknown DNA sequences around a known locus has important applications in molecular genetics, specifically in genomic walking and genome mapping. Several PCR-based methods have been reported to address this issue, but they often involve multiple, time-consuming steps. We have previously described a technique known as restriction site PCR (RS-PCR) that allows sequence acquisi...

Introduction The quality of DNA, obtained by various protocols and subjected to various storage conditions can be judged by comparing the PCR amplification profiles. The most commonly used technique that can be accessed by almost any laboratory is RAPD . Random amplified polymorphic DNA (RAPD) is a PCR based technique for identifying genetic variation. It involves the use of a single arbitrary ...

The purpose of this study was to develop a SNaPshot® assay to simultaneously discriminate between the dystrophic and wild type (wt) alleles in mdx mice. The mdx mouse is an animal model for Duchenne muscular dystrophy (DMD), a severe and fatal muscle wasting disease. To evaluate possible treatments and to carry out genetic studies, it is essential to distinguish between mice that carry the muta...

background inflammatory bowel disease (ibd) is a chronic, relapsing, inflammatory disorder of the gastrointestinal tract that includes two entities, crohn`s disease (cd) and ulcerative colitis (uc). as with other complex diseases, both genetic susceptibility and environmental factors play role in the pathogenesis of these diseases. the tumor necrosis factor α (tnf-α) gene is located in the ibd3...

More sensitive methodologies are necessary to improve strongyloidiasis diagnosis. This study compared the sensitivities of the McMaster modified technique and polymerase chain reaction (PCR) assays, both performed in faecal samples. Lewis rats were subcutaneously infected with 4,000, 400 or 40 infective third-stage larvae, considered as high, moderate or low infection, respectively. Seven days ...

PURPOSE Approximately 95% of patients who are diagnosed with Leber's hereditary optic neuropathy (LHON) have one of three mitochondrial point mutations responsible for the disease, G3460A, G11778A, and T14484C. The purpose of this study was to develop a novel multiplex real-time amplification-refractory mutation system (ARMS) PCR combined with high-resolution melt curves to identify the individ...

Fungi constitute an important group in soil biological diversity and functioning. However, characterization and knowledge of fungal communities is hampered because few primer sets are available to quantify fungal abundance by real-time quantitative PCR (real-time Q-PCR). The aim in this study was to quantify fungal abundance in soils by incorporating, into a real-time Q-PCR using the SYBRGreen®...

The most studied single nucleotide polymorphisms of the VDR gene are Bsm1, Apa1, Taq1 and Fok1. Previously, many approaches have been used to study SNPs in VDR gene including restriction fragment length polymorphism (RFLP), Single Amplification refractory mutation system PCR (Single-ARMS-PCR), and sequencing of the VDR gene. The objective of the study was to develop a multiplex ARMS-PCR system ...

A chromosome walking method was improved in this work. The new method was named as nonspecific primer anchored PCR (NPA-PCR). Nested gene specific primers were designed based on the known region and long random primer using degeneracy oligonucleotides for nonspecific anchoring. Annealing temperatures were varied to control the priming. Target sequences were obtained by PCR with random primer an...