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Review
. 2016 Dec;56(4):394-404.
doi: 10.1007/s12088-016-0606-4. Epub 2016 Jul 9.

High Throughput Sequencing: An Overview of Sequencing Chemistry

Sheetal Ambardar  1 , Rikita Gupta  2 , Deepika Trakroo  2 , Rup Lal  3 , Jyoti Vakhlu  2
Affiliations
Review

High Throughput Sequencing: An Overview of Sequencing Chemistry

Sheetal Ambardar et al. Indian J Microbiol. 2016 Dec.

Abstract

In the present century sequencing is to the DNA science, what gel electrophoresis was to it in the last century. From 1977 to 2016 three generation of the sequencing technologies of various types have been developed. Second and third generation sequencing technologies referred commonly to as next generation sequencing technology, has evolved significantly with increase in sequencing speed, decrease in sequencing cost, since its inception in 2004. GS FLX by 454 Life Sciences/Roche diagnostics, Genome Analyzer, HiSeq, MiSeq and NextSeq by Illumina, Inc., SOLiD by ABI, Ion Torrent by Life Technologies are various type of the sequencing platforms available for second generation sequencing. The platforms available for the third generation sequencing are Helicos™ Genetic Analysis System by SeqLL, LLC, SMRT Sequencing by Pacific Biosciences, Nanopore sequencing by Oxford Nanopore's, Complete Genomics by Beijing Genomics Institute and GnuBIO by BioRad, to name few. The present article is an overview of the principle and the sequencing chemistry of these high throughput sequencing technologies along with brief comparison of various types of sequencing platforms available.

Keywords: High throughput sequencing; NGS; Second generation sequencing; Sequencing platforms; Third generation sequencing.

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Figures

Fig. 1
Fig. 1
Overview of the second and third generation sequencing technologies
Fig. 2
Fig. 2
Diagrammatic representation of enzymatic reactions involved in Pyrosequencing a during complementary nucleotide incorporation and b when nucleotide is not incorporated
Fig. 3
Fig. 3
Diagrammatic representation of Sequencing by reversible termination a during complementary nucleotide incorporation and b when nucleotide is not incorporated
Fig. 4
Fig. 4
Diagrammatic representation of pH change involved in Sequencing by detection of hydrogen ions a during complementary nucleotide incorporation and b when nucleotide is not incorporated
Fig. 5
Fig. 5
Diagrammatic representation of enzymatic reactions involved in Sequencing by hybridization and ligation
Fig. 6
Fig. 6
Diagrammatic representation of single molecule real time (SMRT) sequencing in Zero-mode waveguide a during complementary nucleotide incorporation and b when nucleotide is not incorporated
Fig. 7
Fig. 7
Diagrammatic representation of Helicos single molecule sequencing a during complementary nucleotide incorporation and b when nucleotide is not incorporated
Fig. 8
Fig. 8
Diagrammatic representation of a nanopore sequencing during base calling and b no base case calling
See this image and copyright information in PMC

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