Emulsion PCR: Techniques and applications

Rashmi Kanagal-Shamanna

doi:10.1007/978-1-4939-3360-0_4

Emulsion PCR: Techniques and applications

Rashmi Kanagal-Shamanna

Hematopathology

Research output: Chapter in Book/Report/Conference proceeding › Chapter

12 Scopus citations

Abstract

Emulsion PCR (EmPCR) is a commonly employed method for template amplifi cation in multiple NGSbased sequencing platforms. The basic principle of emPCR is dilution and compartmentalization of template molecules in water droplets in a water-in-oil emulsion. Ideally, the dilution is to a degree where each droplet contains a single template molecule and functions as a micro-PCR reactor. Here, we discuss the basic principles, advantages, and challenges of applications of emPCR in clinical testing. We describe the methods of preparation and enrichment of template-positive Ion PGM™ Template OT2 200 Ion Sphere™ Particles (ISPs) on the Ion Personal Genome Machine ® (PGM™) System. For routine clinical testing, following library generation, we employ the automated Ion OneTouch™ System that includes the Ion OneTouch™ 2 and the Ion OneTouch™ ES instruments for template generation and enrichment of template- positive ISPs, respectively.

Original language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	33-42
Number of pages	10
DOIs	https://doi.org/10.1007/978-1-4939-3360-0_4
State	Published - Feb 1 2016

Publication series

Name	Methods in Molecular Biology
Volume	1392
ISSN (Print)	1064-3745

Keywords

Emulsion PCR
Ion PGM™
Ion sphere™ particles (ISPs)

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-4939-3360-0_4

Cite this

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KW - Ion sphere™ particles (ISPs)

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Emulsion PCR: Techniques and applications

Abstract

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Keywords

ASJC Scopus subject areas

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