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桥式PCR,一种简易连接DNA标签序列的方法
引用本文:毛建平,王全会,周颖,方静,崔玉芳.桥式PCR,一种简易连接DNA标签序列的方法[J].中国生物工程杂志,2009,29(11):66-69.
作者姓名:毛建平  王全会  周颖  方静  崔玉芳
作者单位:北京放射医学研究所 北京 100850
基金项目:科技重大专项,国家自然科学基金 
摘    要:MAST方法采用人工文库的DNA标签序列鉴定mRNA的可接近位点。大量的标签序列通过扩增和克隆测序达到阐明mRNA结合位点图。设计了单一引物的PCR,其引物在标签序列两端结合搭桥,在扩增中DNA标签序列在搭桥引物的作用下进行连接,连接的标签序列再克隆和测序。十几条这样的连接产物包含了上千条标签序列。该PCR方法简单、高效以用于高通量的方式对标签序列测序。

关 键 词:生物反应器  小鼠胚胎干细胞  拟胚体  心肌细胞  分化  
收稿时间:2009-08-03
修稿时间:2009-09-01

Bridge PCR, An Easy Way for Concatemerizing DNA Tags
MA Jian-ping,WANG Quan-hui,ZHOU Ying,FANG Jing,CUI Yu-fang.Bridge PCR, An Easy Way for Concatemerizing DNA Tags[J].China Biotechnology,2009,29(11):66-69.
Authors:MA Jian-ping  WANG Quan-hui  ZHOU Ying  FANG Jing  CUI Yu-fang
Institution:Institute of Radiation Medicine,Beijing 100850, China
Abstract:In MAST (mRNA accessible site tagging) , the DNA tags from synthesized library were employed for identifying mRNA accessible sites. A large number of tags were amplified and subcloned for sequencing to verify mRNA binding profiles. A PCR was designed by using one primer which bridges over the tag terminal sequences. In PCR reaction DNA tag fragments were concatemerized by a bridge primer in reaction cycles. The concatemerized tag fragments were subcloned and sequenced. Dozens of the concatemerized sequences contained thousands tags. The PCR was a simple, effective way which for sequencing tags in a high through put manner.
Keywords:Bioreactor  Mouse embryonic stem cells  Embryoid body  Cardiomyocytes  Differentiation
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