香雪兰花瓣总RNA的提取和cDNA文库的构建  

A method for extraction of total RNA from flowers of freesia and cDNA library construction

作　　者：付杨[1] 高翔[1] 敖曼[1] 王钦美[1] 王丽[1] 

机构地区：[1]东北师范大学遗传与细胞研究所,吉林长春130024

出　　处：《东北师大学报（自然科学版）》2008年第3期118-121,共4页Journal of Northeast Normal University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(30570170);长春市科技发展计划项目(2006258)

摘　　要：以不同开放时间的红色香雪兰的花瓣为材料,利用CTAB裂解、LiCl沉淀的方法提取出高质量的总RNA,经纯化mRNA后,合成双链cDNA,加上EcoRⅠ和XhoⅠ接头,用胶回收的方法将500 bp以上的cDNA回收并连在载体上,获得了重组率为89%,滴度为4.8×105pfu/mL的香雪兰花瓣质粒cDNA文库,为研究香雪兰花香和花色的相关基因奠定了基础.A suitable method for extraction of high quality RNA from freesia＇ s flowers was establish by added CTAB into the extract solution and LiCL to deposit. Separated mRNA from the total RNA. After doublestrand cDNAs synthesis, EcoR Ⅰ and Xho Ⅰ adaptor were added to blunted eDNA. The fragments longer than 500 bp were ligated into the pBluescript Ⅱ XR Predigested Vector. A primery library of 4.8 × 10^5 pfu/mL was obtained. The recombinant efficiency was 89 %. This constracted eDNA library provided an essential resource for future functional genomic analysis of freesia.

关 键 词：香雪兰 总RNA提取 CDNA文库 

分 类 号：Q943.2[生物学—植物学]