一种单链古代DNA文库制备方法。

A Method for Single-Stranded Ancient DNA Library Preparation.

作者信息

Gansauge Marie-Theres, Meyer Matthias

机构信息

Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany.

出版信息

Methods Mol Biol. 2019;1963:75-83. doi: 10.1007/978-1-4939-9176-1_9.

DOI:10.1007/978-1-4939-9176-1_9

PMID:30875046

Abstract

Genomic library preparation from highly degraded DNA is more efficient when library molecules are prepared separately from the complementary strands of DNA fragments. We describe a protocol in which libraries are constructed from single DNA strands in a three-step procedure: single-stranded ligation of the first adapter with T4 DNA ligase in the presence of a splinter oligonucleotide, copying of the DNA strand with a proofreading polymerase, and blunt-end ligation of the second double-stranded adapter with T4 DNA ligase.

摘要

当文库分子从DNA片段的互补链中单独制备时，从高度降解的DNA制备基因组文库效率更高。我们描述了一种在三步过程中从单链DNA构建文库的方案：在分裂寡核苷酸存在下用T4 DNA连接酶将第一个接头进行单链连接，用校对聚合酶复制DNA链，并用T4 DNA连接酶将第二个双链接头进行平端连接。