Liquid drop of DNA libraries reveals total genome information

Terekhov Stanislav S.; Eliseev Igor E.; Ovchinnikova Leyla A.Kabilov Marsel R.Prjibelski Andrey D.Tupikin Alexey E.Smirnov Ivan VBelogurov Alexey A. Jr.Severinov Konstantin VLomakin Yakov A.Altman SidneyGabibov Alexander G.

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Liquid drop of DNA libraries reveals total genome information

机译：Liquid drop of DNA libraries reveals total genome information

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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.

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《Proceedings of the National Academy of Sciences of the United States of America.》 |2020年第44期|27300-27306|共7页
作者
Terekhov Stanislav S.; Eliseev Igor E.; Ovchinnikova Leyla A.Kabilov Marsel R.Prjibelski Andrey D.Tupikin Alexey E.Smirnov Ivan VBelogurov Alexey A. Jr.Severinov Konstantin VLomakin Yakov A.Altman SidneyGabibov Alexander G.;
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作者单位

Shemyakin Ovchinnikov Inst Bioorgan Chem,Russian Acad Sci;

Nanobiotechnol Lab,Alferov Univ;

Inst Chem Biol & Fundamental Med,Russian Acad SciCtr Algorithm Biotechnol,St Petersburg State UnivDept Chem,Lomonosov Moscow State UnivInst Mol Genet,Russian Acad SciDept Mol Cellular & Dev Biol,Yale Univ;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种英语
中图分类自然科学总论;
关键词
quasi single-molecule amplification; uniform distribution of amplicons; diversity degeneration; emulsion PCR modeling; template mispairing; DIRECTED EVOLUTION; EMULSION PCR; DIGITAL PCR; GENERATION;

Liquid drop of DNA libraries reveals total genome information

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