发明授权
- 专利标题: Method for simulataneous identification of differentially expresed mRNAS and measurement of relative concentrations
- 专利标题(中): 用于模拟鉴别差异表达mRNAs的方法和相对浓度的测量
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申请号: US544577申请日: 1995-10-17
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公开(公告)号: US5807680A公开(公告)日: 1998-09-15
- 发明人: J. Gregor Sutcliffe , Mark G. Erlander
- 申请人: J. Gregor Sutcliffe , Mark G. Erlander
- 申请人地址: CA La Jolla
- 专利权人: The Scripps Research Institute
- 当前专利权人: The Scripps Research Institute
- 当前专利权人地址: CA La Jolla
- 主分类号: C12N15/09
- IPC分类号: C12N15/09 ; C07H21/04 ; C12N1/21 ; C12N15/10 ; C12P19/34 ; C12P21/02 ; C12Q1/68 ; C12Q1/70 ; C12R1/19 ; G01N33/48
摘要:
An improved method for the simultaneous sequence-specific identification of mRNAs in a mRNA population allows the visualization of nearly every mRNA expressed by a tissue as a distinct band on a gel whose intensity corresponds roughly to the concentration of the mRNA. In general, the method comprises the formation of cDNA using anchor primers to fix a 3'-endpoint, producing cloned inserts from the cDNA in a vector containing a bacteriophage-specific promoter for subsequent RNA synthesis, generating linearized fragments of the cloned inserts, preparing cRNA, transcribing cDNA from the cRNA using a set of primers, and performing PCR using a 3'-primer whose sequence is derived from the vector and a set of 5'-primers that is derived from the primers used for transcription of cDNA from cRNA. The method can identify changes in expression of mRNA associated with the administration of drugs or with physiological or pathological conditions.
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