福建农业学报2012，Vol.27Issue(11)：1178-1182,5.
利用改进的SMART法构建花生种皮全长cDNA文库
Establishment of Full-length Peanut Testa cDNA Library by Using Modified SMART
摘要
Abstract
SMART is one of the most important methods for constructing a full-length cDNA library. In order to discover new genes from the peanut testa, the SMART method published previously was modified to improve its efficiency, reduce cost, simplify operation, and minimize test time. Using the optimized protocol, a full-length cDNA library for peanut testa was established. The new method produced an estimated titer of the amplified cDNA library as high as 3. 84×10~9cfu o mL-1, which was increased from 1. 23×10~6 cfu o mL-1 , with a recombinant rate of 93. 3%. Furthermore, approximately 70% of the inserts, varying from 1. 0 to 2. 0 kb with an average size of about 1 100 bp, were obtained. The sequence analysis indicated that the full-length cDNA proportion was as high as 55%. It was concluded that the library was of desirable quality for full-length screening and cloning of the target genes.关键词
SMART法/全长cDNA文库/花生种皮Key words
SMART/ full-length cDNA libraty/ peanut testa分类
农业科技引用本文复制引用
陈华,姜宝杰,张冲,曾建斌,邓烨,庄伟建..利用改进的SMART法构建花生种皮全长cDNA文库[J].福建农业学报,2012,27(11):1178-1182,5.基金项目
科技部国际合作项目(2008DFA31150) （2008DFA31150）
