期刊论文

中文

分子植物育种

1672-416X

2010

8

4

800-803

10.3969/mpb.008.000800

国家科技支撑计划（2006BAD19B0901） 国家高技术研究发展计划重点项目（2006AA100109）共同资助

本校为第一机构

本实验比较了3 种RNA 提取方法提取单叶省藤的茎、雄花序和根的结果, 改良CT 法和Trizol法提取的RNA 无明显降解、完整性好, 均可用于cDNA 文库构建, 而异硫氰酸胍法提取的RNA 降解严重; 考虑到改良CT 法成本较低, 故推荐为首选方法. 将茎、雄花序和根三种组织的RNA 等量混合后, 利用 SMART 策略构建了全长cDNA 文库; 通过涂平板培养后的菌落计数, 测得初库滴度为2.91×10~5 cfu/μL; 从培养平板上随机挑取16 个菌落进行PCR 检测, 文库重组率为87.5%, 插入片段平均长度为1.5 kb 左右. 所建文库的质量可靠, 为下一步的EST 测序和候选基因发现提供了材料基础

In this research,by comparing the results among three procedures for isolating total RNA from stem,male inflorescence and root of Calamus simplicifolius,we found that modified CTAB method and Trizol method were better than guanidnium isothiocyanate method on yield and quality of RNA from all the three tisues.As modified CTAB method was more cost-effective than the Trizol,it was recommended as the sound choice of RNA isolation.Then,we constructed a full-length cDNA library in accordance with SMART strategy and by using a mixture of RNA of the th...