Hall Bradley, Arshad Seyed, Seo Kyunghyun, Bowman Catherine, Corley Meredith, Jhaveri Sulay D, Ellington Andrew D
Department of Chemistry and Biochemistry, University of Texas, Austin, Texas, USA.
Curr Protoc Nucleic Acid Chem. 2010 Mar;Chapter 9:Unit 9.3.1-27. doi: 10.1002/0471142700.nc0903s40.
This unit describes the selection of aptamers from a pool of single-stranded RNA by binding to a protein target. Aptamers generated from this selection experiment can potentially act as protein function inhibitors, and may find applications as therapeutic or diagnostic reagents. A pool of dsDNA is used to generate an ssRNA pool, which is mixed with the protein target. Bound complexes are separated from unbound reagents by filtration, and the RNA:protein complexes are amplified by a combination of reverse transcription, PCR, and in vitro transcription.
本单元描述了通过与蛋白质靶标结合从单链RNA库中筛选适配体的过程。从该筛选实验中产生的适配体有可能作为蛋白质功能抑制剂，并可能作为治疗或诊断试剂得到应用。双链DNA库用于生成单链RNA库，该单链RNA库与蛋白质靶标混合。通过过滤将结合的复合物与未结合的试剂分离，然后通过逆转录、聚合酶链反应（PCR）和体外转录相结合的方式扩增RNA：蛋白质复合物。
