发明授权
US6083727A Methods and compositions for producing 5' enriched cDNA libraries
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用于产生5'富集的cDNA文库的方法和组合物
- 专利标题: Methods and compositions for producing 5' enriched cDNA libraries
- 专利标题(中): 用于产生5'富集的cDNA文库的方法和组合物
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申请号: US343945申请日: 1999-06-30
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公开(公告)号: US6083727A公开(公告)日: 2000-07-04
- 发明人: Karl Guegler , Ruoying Tan , Michael J. Rose
- 申请人: Karl Guegler , Ruoying Tan , Michael J. Rose
- 申请人地址: CA Palo Alto
- 专利权人: Incyte Pharmaceuticals Inc.
- 当前专利权人: Incyte Pharmaceuticals Inc.
- 当前专利权人地址: CA Palo Alto
- 主分类号: C12N15/09
- IPC分类号: C12N15/09 ; C12N15/10 ; C12Q1/68 ; C07H21/02 ; C07H21/04 ; C12P19/34
摘要:
Methods are provided for preparing a double-stranded cDNA corresponding to the 5' end of an mRNA. In the subject methods, an mRNA is first contacted with an oligo dT primer under first strand cDNA synthesis conditions. Next, the resultant hybrid is contacted with a random primer under first strand cDNA synthesis conditions, such that a cDNA complementary to the 5' end of the mRNA is produced. The resultant hybrid molecule is converted to two different double-stranded cDNAs, a first cDNA comprising the oligo dT primer and a second cDNA lacking the oligo dT primer. The two double-stranded cDNAs are then separated from each other. The subject methods find use in a variety of applications, and find particular use in the synthesis of 5' enriched cDNA libraries. Also provided are cDNA libraries produced by the subject methods, as well as kits for performing the subject methods.
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