Genetic Systems, Life Technologies Inc., South San Francisco, CA 94080, USA.
Proc Natl Acad Sci U S A. 2013 Aug 27;110(35):14320-3. doi: 10.1073/pnas.1311334110. Epub 2013 Aug 12.
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.
我们报告了一种用于下一代测序应用的固定化单克隆模板的生成方法。我们的等温扩增方法基于使用一对低熔点温度（Tm）固相亲聚物引物和低 Tm 溶液相引物的模板行走机制。该方法可以在单个下一代测序流片通道中生成超过十亿个亚微米大小的菌落。还展示了一种替代的配对末端测序方法，该方法使用链间 DNA 光交联将原始模板的互补链共价连接到固体表面。
