Next Generation Sequencing

This Review focuses on commercially available technologies from Roche/454, Illumina/Solexa, Life/APG and Helicos BioSciences, the Polonator instrument and the near-term technology of Pacific Biosciences,

Here, I present a technical review of template preparation, sequencing and imaging, genome alignment and assembly, and current NGS platform performance to provide guidance on how these technologies work and how they may be applied to important biological questions. ​

Sequencing technologies include a number of methods
that are grouped broadly as template preparation,
sequencing and imaging, and data analysis.

• There are two methods used in preparing templates for NGS reactions: clonally amplified templates originating from single DNA molecules, and single DNA molecule templates

Clonally amplified templates. most imaging systems have
not been designed to detect single fluorescent events, so
amplified templates are required. The two most common
methods are emulsion PCR (emPCR)9 and solid-phase
amplification

emulsion PCR (emPCR)

solid-phase amplification​