Knudsen Scott M, Robertson Michael P, Ellington Andrew D
University of Texas, Austin, Texas, USA.
Curr Protoc Nucleic Acid Chem. 2002 Feb;Chapter 9:Unit 9.6. doi: 10.1002/0471142700.nc0906s07.
The use of modified nucleotides in an RNA or DNA pool to be used for in vitro selection offers many potential advantages, such as the increased stability of the selected nucleic acid against nuclease degradation. This unit provides useful information and protocols for in vitro selection using modified nucleotides. It includes a discussion of when to use modified nucleotides; protocols for preparing a modified RNA pool and verifying its suitability for in vitro selection; and protocols for selecting and amplifying a functionally enriched pool.
在用于体外筛选的RNA或DNA文库中使用修饰核苷酸具有许多潜在优势,例如所选核酸对核酸酶降解的稳定性增强。本单元提供了关于使用修饰核苷酸进行体外筛选的有用信息和实验方案。它包括何时使用修饰核苷酸的讨论;制备修饰RNA文库并验证其是否适合体外筛选的实验方案;以及筛选和扩增功能富集文库的实验方案。