FENG Pei-chun,CUI Yi-hui,ZHANG Zi-feng,et al.cDNA PCR Library of mRNA From Dendrobium candidum Leaves Constructed Using RACE Technique[J].Northern Horticulture,2014,38(11):85-88.
通过RACE技术构建铁皮石斛叶mRNA的cDNA PCR 文库
- Title:
- cDNA PCR Library of mRNA From Dendrobium candidum Leaves Constructed Using RACE Technique
- 文章编号:
- 1001-0009(2014)11-0085-04
- 关键词:
- cDNA PCR文库; RACE; 逆转录; PCR
- Keywords:
- cDNA PCR library; RACE; reverse transcription; PCR
- 分类号:
- R 931. 71
- 文献标志码:
- A
- 摘要:
- 基于RACE技术，以铁皮石斛叶片总RNA为起始材料，通过逆转录和末端转移构建出两端含有特定序列的cDNA。然后以连接有特定序列的引物进行cDNA的PCR扩增，从而构建出mRNA的cDNA PCR文库。结果表明：cDNA PCR文库的PCR效果明显高于逆转录产物的PCR效果，该方法构建的cDNA PCR文库能使cDNA放大100倍以上；以5 pg的总RNA为起始材料构建cDNA PCR文库仍可得到良好的PCR扩增效果；该研究的cDNA PCR文库构建方法费用较低，操作简单，为克隆微量表达的基因提供了技术基础。
- Abstract:
- Based on RACE technique,total RNA from Dendrobium candidum leaves was subjected to reverse transcription and terminal deoxynucleotidyl transfer resulting the cDNA which contained the specific sequences in two ends.The obtained cDNA with the specific sequences was amplified using PCR with primers which matched the specific sequences to establish cDNA PCR library.The results showed that PCR amplification of the cDNA PCR library was more efficient significantly than that of cDNA.The quantity of cDNA was magnified more than one hundred times by the constructed cDNA PCR library.By the cDNA PCR library constructed using 5 pg total RNA as material,the satisfactory PCR amplification results can be obtained.This construction method of the cDNA PCR library was time and labor saving.This method would provided the technical basis for cloning genes of low expression.
参考文献/References:
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[3] 李晶泉,袁晓东,汤敏谦.微量 RNA 的 cDNA PCR 文库的构建 [J].遗传,2001,23(2):147-150.
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备注/Memo
第一作者简介：冯沛春(1989-)，男，硕士，研究方向为植物资源与分子生物学。E-mail：peichunfeng@163.com.
责任作者：王万军(1962-)，男，博士，教授，硕士生导师，研究方向为植物资源与生物技术。E-mail：wanjunwang@home.swjtu.edu.cn.
基金项目：国家自然科学基金资助项目(31271302)。
收稿日期：2014-01-20