The Generation Sequencing ( Ngs ) Technologies

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Introduction:

Since the past decade, the next generation sequencing (NGS) technologies have developed the whole genome analysis field (1). The rapid development in NGS methods provide large sequencing data for each patient (3). Next generation sequencing technology enables generation of millions of DNA fragments in parallel which lead to produce a new era in medical genomics (2). These technologies have been involved in several studies including; RNA sequencing, bisulfite sequencing, DNA target, resequencing, Chromatin Immunoprecipitation sequencing, MethylCap-seq and others (3). NGS have enumerous developed platforms such as illumina/Solexa, SOLID, 454 pyrosequencing, Pacific Bio, and Ion Torrent Sequencing (5). In 2009, a study showed that NGS technologies have been applied to invistigate mRNA expression, genome sequence variations, post transcriptional and post translational (5). -----------

RNA Sequencing:

RNA-Seq is technique that allows to quantify gene expession patterns for RNA profiling dependent on NGS (4). Before RNA-Seq methodology has been used, scientists were using Microarray technique for gene expression study (4). RNA sequencing framework enables to examine the presence of all RNAs in a study sample, differentiating their sequences as well as determing their abundances simultaneously. RNA-Seq method takes place to any of many various techniques of NGS applied for gaining whole transcriptome profiles of RNA that can be explored in cell, tissue and other multicellular organisms (5). In addition, all of NGS technologies can be applied for RNA sequencing method. Each of these technologies or aggregation of them have been used as distict RNA-Seq technologies. Despite they share the same concept to produce RNA seq...

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...matics strategies exist to measure gene expression through different examination samples in the RNA-Seq method. There are several methods are emplyed for calculating differential gene such as Reads per Kilobase of Exon per Million of Mapped reads, upper-quartile and Trimmed Means of M Values. To quantify relatively significant differential gene expression, Empirical Bayesian method, Bayesian methods and Negative binominal distribution stategies can be performed (5).

However, using bioinformatics in RNA-Seq can analyze Single Nucletide Ploymorphism for example; Sequence Alignment Map is a general desgin for collecting and saving the alignmets of large nucleotide sequence which is then utilized for SNP analyses. The following steps for determing SNPs is dependent on converting SAM to Binary Variant Call Format file and then BFC file to Variant Call Format file (5).

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