发明名称 |
CONSTRUCTION OF UNI-DIRECTIONALLY CLONED cDNA LIBRARIES FROM MESSENGER RNA FOR IMPROVED 3'END DNA SEQUENCING |
摘要 |
Methods are provided for preparing cDNA corresponding to a mRNA. In the subject methods, a mRNA is first contacted with a mixture of primers under first strand cDNA synthesis conditions. The primer mixture contains primers that have at least 10 contiguous deoxythymidines, a double stranded restriction enzyme recognition sequence near one end and a non-polyA-complementary region near the other end, where the non-polyA-complementary region is -VV, -VTV, -VTTV, -VTTTV, and -VVVVV. The resultant cDNA is modified such that the polyT tail is substantially removed. The modified cDNA is then ligated into a vector. The subject methods find use in a variety of applications, and find particular use in the sequencing of DNA and in the synthesis of cDNA libraries. |
申请公布号 |
WO0179549(A3) |
申请公布日期 |
2002.05.16 |
申请号 |
WO2001US11211 |
申请日期 |
2001.04.05 |
申请人 |
INCYTE PHARMACEUTICALS, INC. |
发明人 |
FU, GLENN, K.;STARNES, STEVEN;STUVE, LAURA, L. |
分类号 |
C12N15/10 |
主分类号 |
C12N15/10 |
代理机构 |
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地址 |
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