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Updated: May 8, 2026

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Published on: August 29, 2014
Isothermal amplification method for next-generation sequencing
Zhaochun Ma1, Raymond W Lee, Bin Li
1Genetic Systems, Life Technologies Inc., South San Francisco, CA 94080, USA. zhaochun.ma@lifetech.com
Abstract:
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.
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