发明名称 |
Method for making full-length cDNA libraries |
摘要 |
Disclosed is a method for making full-length CDNA libraries, which is for making libraries of cDNAs having a length corresponding to a full-length of mRNAs and comprises the following steps of; binding a tag molecule to a diol structure present in 5' Cap (7MeGpppN) sites of mRNAs, forming RNA-DNA hybrids by reverse transcription using primers such as oligo dT and the mRNAs connected with the tag molecule as templates, and separating RNA-DNA hybrids carrying a DNA corresponding to a full-length of mRNAs from the RNA-DNA hybrids formed above by using function of the tag molecule. To obtain mRNA connected with a tag molecule, the diol structure present in 5' Cap site of mRNA is subjected to a ring-open reaction by oxidation with sodium periodate to form a dialdehyde and the dialdehyde is reacted with a tag molecule having a hydrazine terminus. According to the present invention, there are provided a novel method capable of efficiently labeling 5' Cap site and a method for making full-length cDNA libraries utilizing the labeling method.
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申请公布号 |
US6174669(B1) |
申请公布日期 |
2001.01.16 |
申请号 |
US19960752540 |
申请日期 |
1996.11.20 |
申请人 |
THE INSTITUTE OF PHYSICAL AND CHEMICAL RESEARCH |
发明人 |
HAYASHIZAKI YOSHIHIDE;CARNINCI PIERO;SCHNEIDER CLAUDIO |
分类号 |
C12N15/09;C07B61/00;C07H21/00;C07H21/02;C07H21/04;C12N15/10;(IPC1-7):C12Q1/68 |
主分类号 |
C12N15/09 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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