Liquid drop of DNA libraries reveals total genome information

Stanislav S. Terekhov, Igor E. Eliseev, Leyla A. Ovchinnikova, Marsel R. Kabilov, Andrey D. Prjibelski, Alexey E. Tupikin, Ivan V. Smirnov, Alexey A. Belogurov, Konstantin V. Severinov, Yakov A. Lomakin, Sidney Altman, Alexander G. Gabibov

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Conventional “bulk” PCR often yields inefficient and nonuniform amplification of complex templates in DNA libraries, introducing unwanted biases. Amplification of single DNA molecules encapsulated in a myriad of emulsion droplets (emulsion PCR, ePCR) allows the mitigation of this problem. Different ePCR regimes were experimentally analyzed to identify the most robust techniques for enhanced amplification of DNA libraries. A phenomenological mathematical model that forms an essential basis for optimal use of ePCR for library amplification was developed. A detailed description by high-throughput sequencing of amplified DNA-encoded libraries highlights the principal advantages of ePCR over bulk PCR. ePCR outperforms PCR, reduces gross DNA errors, and provides a more uniform distribution of the amplified sequences. The quasi single-molecule amplification achieved via ePCR represents the fundamental requirement in case of complex DNA templates being prone to diversity degeneration and provides a way to preserve the quality of DNA libraries.

Original languageEnglish (US)
Pages (from-to)27300-27306
Number of pages7
JournalProceedings of the National Academy of Sciences of the United States of America
Volume117
Issue number44
DOIs
StatePublished - Nov 3 2020

Keywords

  • Diversity degeneration
  • Emulsion PCR modeling
  • Quasi single-molecule amplification
  • Template mispairing
  • Uniform distribution of amplicons

ASJC Scopus subject areas

  • General

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