发明名称 |
5'-thio phosphate directed ligation of oligonucleotides and use in detection of single nucleotide polymorphisms |
摘要 |
The present invention provides a novel method for ligation of oligonucleotides containing 5'-phosphorothioates on complementary templates by the action of DNA ligases. This reaction is readily applied to the synthesis of a single stranded circular DNA containing a phosphorothioate linkage at the site of ligation junction. The efficiency of 5'-phosphorothioate directed ligation reaction by ATP dependent DNA ligase reaction is similar to conventional 5'-phosphate ligation. The utility of enzymatic ligation in probing specific sequences of DNA is also described. The present invention also provides a novel non-enzymatic ligation of 5'-phosphorothioates that has been applied to the synthesis of single strand phosphorothioate and phosphate circular DNA. A process for detecting the presence of a mismatch in an otherwise complementary pair of oligonucleotides is disclosed using an enzyme-based technique which shows the presence of a mismatch by failing to form a ligated single stranded DNA circle that can optionally be amplified using standard methods of rolling circle amplification.
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申请公布号 |
US6635425(B2) |
申请公布日期 |
2003.10.21 |
申请号 |
US20010910372 |
申请日期 |
2001.07.20 |
申请人 |
MOLECULAR STAGING, INC. |
发明人 |
BANDARU RAJANIKANTH;KUMAR GYANENDRA |
分类号 |
C12Q1/68;(IPC1-7):C12Q1/68;C12P19/34;C07H21/02 |
主分类号 |
C12Q1/68 |
代理机构 |
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主权项 |
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地址 |
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