dna sequence determination is a tremendous human achievement. dna sequencing includes several methods and technologies in use for determining the order of the nucleotide bases (adenine, guanine, cytosine, and thymine) in a molecule of dna. knowledge of dna sequences has become indispensable for basic biological research, other research branches utilizing dna sequencing, and in numerous applied ...

Accurate duplication of parental DNA is a fundamental biological process, conserved in function across all life forms. All organisms depend on DNA polymerases for genome replication and maintenance. DNA polymerases also play central roles in modern molecular biology and biotechnology, enabling techniques including DNA cloning, the polymerase chain reaction (PCR), DNA sequencing, single nucleoti...

We are developing novel methods to simultaneously analyze candidate genes or regions from multiple samples in a single experiment rapidly and cost-effectively. One method, we call Reflex, provides an elegant approach to sample preparation for long-range PCR (LR-PCR) amplicons. Current methods use LR-PCR for target enrichment then use random fragmentation of each sample followed by ligation of D...

BACKGROUND For next generation DNA sequencing, we have developed a rapid and simple approach for preparing DNA libraries of targeted DNA content. Current protocols for preparing DNA for next-generation targeted sequencing are labor-intensive, require large amounts of starting material, and are prone to artifacts that result from necessary PCR amplification of sequencing libraries. Typically, sa...

The purpose of this experiment was to implement and evaluate the effectiveness of a next-generation sequencing-based method for DNA methylation analysis in porcine embryonic samples. Fourteen discrete genomic regions were amplified by PCR using bisulfite-converted genomic DNA derived from day 14 in vivo-derived (IVV) and parthenogenetic (PA) porcine embryos as template DNA. Resulting PCR produc...

Background: Mucopolysaccharidosis type-VI (MPS-VI), which is inherited as an autosomal recessive trait, results from the deficiency of N-acetylgalactosamine 4-sulfatase (arylsulfatase B) activity and the lysosomal accumulation of dermatan sulfate. In this study, ARSB mutation analysis was performed on three unrelated patients who were originally from the West Azerbaijan province of Iran. Method...

We have developed an NGS-based deep bisulfite sequencing protocol for the DNA methylation analysis of genomes. This approach allows the rapid and efficient construction of NGS-ready libraries with a large number of PCR products that have been individually amplified from bisulfite-converted DNA. This approach also employs a bioinformatics strategy to sort the raw sequence reads generated from NG...

Aberrant hypo-methylation of DNA is evident in a range of human diseases including cancer and diabetes. Development of sensitive assays capable of detecting traces of un-methylated DNA within methylated samples can be useful in several situations. Here we describe a new approach, fast-COLD-MS-PCR, which amplifies preferentially un-methylated DNA sequences. By employing an appropriate denaturati...

Background Leprosy is one of the neglected tropical diseases (NTD) experiencing relative stagnancy in global number new cases. Objective This study aims to find proportion antimicrobial resistance clinical specimens leprosy patients attending a tertiary care Institute South India. Materials and methods Skin biopsy (31 Nos) were tested by Polymerase chain reaction (PCR) for identification M. lep...

Mutations in the gene encoding filaggrin (FLG) have been identified as cause of ichthyosis vulgaris (IV) and shown to be major predisposing factors for atopic dermatitis. The profilaggrin protein consists 10-12 monomer repeats arranged tandem, which arise from an ultra-long sequence exon 3 comprised near-indistinguishable repeats, demonstrating intragenic copy number variations (CNVs). As such,...