发明申请
- 专利标题: Method for Generating Transcripts
- 专利标题(中): 生成记录的方法
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申请号: US11660952申请日: 2005-09-23
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公开(公告)号: US20070260053A1公开(公告)日: 2007-11-08
- 发明人: Francois Mallet , Guy Oriol , Jean-Philippe Pichon
- 申请人: Francois Mallet , Guy Oriol , Jean-Philippe Pichon
- 优先权: FR0452228 20041001
- 国际申请: PCT/FR05/50775 WO 20050923
- 主分类号: C07H21/04
- IPC分类号: C07H21/04 ; C12N9/00
摘要:
The present invention relates to a method for generating transcripts from: at least one RNA sequence to be amplified comprising a “primer” region and a region of interest, and an amplification primer comprising a promoter region, and a region capable of hybridizing to said “primer” region of the RNA sequence to be amplified, said method being carried out at constant temperature, and comprising the following steps: a) said primer is hybridized with the RNA to be amplified, b) the primer is extended by means of a reverse transcriptase enzymatic activity in order to generate a complementary deoxyribonucleic acid (cDNA) sequence of the RNA to be amplified, c) the RNA to be amplified, hybridized to said cDNA, is cleaved by means of an enzyme that has a ribonuclease H activity, so as to obtain fragments of RNA to be amplified, hybridized to said cDNA, d) the ends of said fragments of RNA to be amplified are extended by means of a reverse transcriptase and strand displacement enzyme, so as to obtain RNA-DNA/DNA hybrids, e) RNA transcripts are obtained from the RNA-DNA/DNA hybrids formed in step d), by means of an enzyme that has an RNA polymerase activity.
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