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
Proc. Natl. Acad. Sci. USA, 2020, 117(44), 27300-27306
https://doi.org/10.1073/pnas.2017138117

Abstract

DNA libraries are predisposed to template mispairing during conventional “bulk” PCR, leading to the loss of unique sequences. The latter is facilitated by the nonuniform distribution of templates frequently observed in DNA libraries. These effects result in a prominent reduction of the original diversity. The encapsulation of DNA repertoires in liquid droplets abolishes the effects of mispairing in DNA libraries. The fundamental advantages of emulsion PCR (ePCR) over bulk PCR are illustrated by deep sequencing and mathematical modeling, which provide the general strategy for ePCR rationalization. The quasi single-molecule ePCR reveals total genetic information by counteracting the degeneration of DNA libraries’ diversity.