Abstract
Emulsion PCR (EmPCR) is a commonly employed method for template amplification in multiple NGS-based 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.
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Kanagal-Shamanna, R. (2016). Emulsion PCR: Techniques and Applications. In: Luthra, R., Singh, R., Patel, K. (eds) Clinical Applications of PCR. Methods in Molecular Biology, vol 1392. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3360-0_4
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DOI: https://doi.org/10.1007/978-1-4939-3360-0_4
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3358-7
Online ISBN: 978-1-4939-3360-0
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