Personal Diagnostics Using DNA-Sequencing

Personal Diagnostics Using DNA-Sequencing

Udayaraja GK
Copyright: © 2016 |Pages: 16
DOI: 10.4018/978-1-4666-8726-4.ch009
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Abstract

DNA sequencing is the process to identification of nucleotides order in genome which developed from very broad history, also it is derived from version of the Sanger biochemistry. SOLiD, 454 and Polonator sequencing based on emulsion PCR to amplify clonal sequencing with in-vitro construction of adaptor-flanked shotgun library, PCR amplified in the context of a water-in-oil emulsion. Solexa technology relies on bridge PCR to amplify clonal sequencing features. At the conclusion of the PCR, each clonal cluster contains ~1,000 copies of a single member of the template library. This chapter focused on next-generation sequencing technologies methods, capabilities and clinical applications of DNA sequencing technologies for researchers in molecular biology and physician scientists. This will also provide the power of these novel genomic tools and methods to use personal diagnostic at molecular level.
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Background

NGS platforms produce a massive amount data (up to terabases) in parallel sequencing method. Often, NGS platforms are classified as second and third generation sequencing technologies. The methods which depends on a PCR step for signal intensification prior to sequencing as next generation sequencing instruments, opposed to single molecule sequencing. Next generation sequencing technology includes the 454 instruments from Roche, the different Illumina platforms and the Life Technologies instruments, i. e. the SOLiD (Sequencing by Oligonucleotide Ligation and Detection) and PacBio RS by Pacific Biosciences. The third generation sequencing instruments like Ion Torrent(PGM=Personal Genome Machine, IonProton) sequencers, MySeq & MsJunior. Next-Generation technology platforms differ in read length, throughput size, data metrics, read depth, etc (Venter et al., 2001; Mardis, 2008).

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