Isothermal amplification method for next-generation sequencing

被引：17

作者：

Ma, Zhaochun ^{[1
]}

Lee, Raymond W. ^{[1
]}

Li, Bin ^{[1
]}

Kenney, Paul ^{[1
]}

Wang, Yufang ^{[1
]}

Erikson, Jonathan ^{[1
]}

Goyal, Swati ^{[1
]}

Lao, Kaiqin ^{[1
]}

机构：

[1] Life Technol Inc, Genet Syst, San Francisco, CA 94080 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2013年 / 110卷 / 35期

关键词：

genome; in situ PCR; forming colonies; DNA breathing; polony; PHOTO-CROSS-LINKING; DNA-MOLECULES; GENOME; COLONIES;

D O I：

10.1073/pnas.1311334110

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

We report an approach for generating immobilized monoclonal templates for next- generation sequencing applications. Our isothermal amplification method is based on a template walking mechanism using a pair of low-melting temperature (Tm) solid-surface homopolymer primers and a low-Tm solution phase primer. The method can generate more than one billion submicrometer-sized colonies in a single lane of a next- generation sequencing flowchip. An alternative paired-end sequencing method using interstrand DNA photo cross-linking to covalently link the complementary strands of the original templates to the solid surface is also demonstrated.

引用

页码：14320 / 14323

页数：4

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