用于体外筛选的DNA文库的设计、合成及扩增

Design, synthesis, and amplification of DNA pools for in vitro selection.

作者信息

Hall Bradley, Micheletti John M, Satya Pooja, Ogle Krystal, Pollard Jack, Ellington Andrew D

机构信息

Department of Chemistry and Biochemistry, University of Texas, Austin, Texas, USA.

出版信息

Curr Protoc Nucleic Acid Chem. 2009 Dec;Chapter 9:Unit 9.2. doi: 10.1002/0471142700.nc0902s39.

DOI:10.1002/0471142700.nc0902s39

PMID:20013786

Abstract

Preparation of a random-sequence DNA pool is presented. The degree of randomization and the length of the random sequence are discussed, as is synthesis of the pool using a DNA synthesizer or via commercial synthesis companies. Purification of a single-stranded pool and conversion to a double-stranded pool are presented as step-by-step protocols. Support protocols describe determination of the complexity and skewing of the pool, and optimization of amplification conditions.

摘要

介绍了随机序列DNA文库的制备方法。讨论了随机化程度和随机序列的长度，以及使用DNA合成仪或通过商业合成公司合成文库的方法。逐步方案介绍了单链文库的纯化和转化为双链文库的方法。支持方案描述了文库复杂性和偏差的测定，以及扩增条件的优化。