Next generation sequencing

الرحیم الرحمن الله بسم

Next Generation Sequencing

By: Ehsan Sepahi

A Timeline of genetics

Sanger and Maxam-Gilbert

Sanger and Maxam-Gilbert

Next Generation Sequencing• The automated Sanger method is considered as

a ‘first-generation’ technology, and newer methodsare referred to as next-generation sequencing (NGS)

• These newer technologies constitute various strategies that rely on a combination of:• template preparation• sequencing and imaging• and genome alignment and assembly

Next Generation Sequencing Platforms

Template preparation• Clonally amplified templates

• emulsion PCR (emPCR) • solid-phase amplification

• Single-molecule templates• primer immobilized• template immobilized• polymerase immobilized

emulsion PCR (emPCR)

solid-phase amplification

primer immobilized

template immobilized

polymerase immobilized

sequencing and imaging• Cyclic reversible termination (Sequencing by

synthesis): Illumina, Helicos BioSciences• Sequencing by ligation: SOLiD• Pyrosequencing: 454• Real-time sequencing: Pasific BioSciences• Ion Semiconductor: Ion Torrent

Sequencing by synthesis

Sequencing by ligation

Pyrosequencing

Real-time sequencing

Ion Semiconductor

Sequencing by synthesis

A Timeline of genetics

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