用于深度测序的小RNA cDNA文库构建。

Construction of small RNA cDNA libraries for deep sequencing.

作者信息

Lu Cheng, Meyers Blake C, Green Pamela J

机构信息

Department of Plant and Soil Sciences, Delaware Biotechnology Institute, University of Delaware, Newark, DE 19711, USA.

出版信息

Methods. 2007 Oct;43(2):110-7. doi: 10.1016/j.ymeth.2007.05.002.

DOI:10.1016/j.ymeth.2007.05.002

PMID:17889797

Abstract

Small RNAs (21-24 nucleotides) including microRNAs (miRNAs) and small interfering RNAs (siRNAs) are potent regulators of gene expression in both plants and animals. Several hundred genes encoding miRNAs and thousands of siRNAs have been experimentally identified by cloning approaches. New sequencing technologies facilitate the identification of these molecules and provide global quantitative expression data in a given biological sample. Here, we describe the methods used in our laboratory to construct small RNA cDNA libraries for high-throughput sequencing using technologies such as MPSS, 454 or SBS.

摘要

包括微小RNA（miRNA）和小干扰RNA（siRNA）在内的小RNA（21 - 24个核苷酸）是植物和动物基因表达的有效调节因子。通过克隆方法已通过实验鉴定出数百个编码miRNA的基因和数千个siRNA。新的测序技术有助于这些分子的鉴定，并在给定的生物样品中提供全局定量表达数据。在这里，我们描述了我们实验室中使用的方法，以使用MPSS、454或SBS等技术构建用于高通量测序的小RNA cDNA文库。