一种改良的扩增cDNA 5′末端的方法

园艺学报 ›› 2008, Vol. 35 ›› Issue (10): 1533-1538.

一种改良的扩增cDNA 5′末端的方法

夏瑞^1,3,陆旺金²,李建国^1*

（¹华南农业大学中国荔枝研究中心，广州 510642; ²广东省果蔬保鲜重点实验室广州 510642; ³广东农业科学院果树研究所，广州510640 ）

收稿日期:2008-05-29 修回日期:2008-09-12 出版日期:2008-10-25 发布日期:2008-10-25
通讯作者: 李建国

A Modified Method of Amplifying the 5′-end cDNA Sequence

XIA Rui^1,3;LU Wang-jin2;and LI Jian-guo^1*

(¹China Litchi Research Center, South China Agricultural University, Guangzhou 510642, China; ²Guangdong Provincial Key Lab of Post-harvest Science of Fruit and Vegetables, Guangzhou 510642, China; ³Institute of Fruit Research, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China)

Received:2008-05-29 Revised:2008-09-12 Online:2008-10-25 Published:2008-10-25
Contact: LI Jian-guo

摘要/Abstract

摘要： 介绍一种改良的扩增基因5′cDNA序列的方法（改良TdT加尾扩增法），该方法以龙眼为材料采用了TdT（末端脱氧核苷酸转移酶）加尾、锚定PCR、巢式PCR、降落PCR等多种策略。通过与常规方法（用TaKaRa试剂盒）的对比试验，发现该方法原理简单，操作性强；扩增特异性高，结果真实可靠；同时还具有快速低廉的特点，适合于在一般实验室中推广使用。

关键词: TdT, RACE, 降落PCR, Dl-Exp1

Abstract:

In order to obtain the full-length sequence of genes more efficiently, a modified method of amplifying the 5′ cDNA sequence, named as modified TDT tailing amplified method, was designed in this experiment. Many strategies are used in this method including TdT tailing, anchored PCR, nested PCR, touchdown PCR. Compared with a regular method using a kit (5′-Full RACE Core Set, TaKaRa), the modified method has many advantages, such as simple principle, easy operability, high amplified specificity, and reliable results. Moreover, it costs less money and time. Therefore, it is worth popularizing in ordinary laboratories.

Key words: TdT, RACE, touchdown PCR, Dl-Exp1

中图分类号:

Q 943.2

夏瑞;陆旺金;李建国. 一种改良的扩增cDNA 5′末端的方法[J]. 园艺学报, 2008, 35(10): 1533-1538.

XIA Rui;LU Wang-jin;and LI Jian-guo. A Modified Method of Amplifying the 5′-end cDNA Sequence [J]. ACTA HORTICULTURAE SINICA, 2008, 35(10): 1533-1538.

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