申请人:Naxcor
公开号:US20030039961A1
公开(公告)日:2003-02-27
Methods and compositions are provided for detecting nucleic acid sequences. In particular, pairs of probes are employed, where the pair defines a substantially contiguous sequence on a target nucleic acid. Each of the pairs has a side chain which forms a stem of the two side chains which non-covalently binds and is capable of forming a cross-link upon activation, when the probes and sample nucleic acid are base paired. Cross-linking of the stems when unbound to complementary DNA is inhibited. Each of the nucleic acids is initially present as single stranded nucleic acid to allow for base pairing, so that the probes bind to homologous target nucleic acid. The assay mixture is activated to provide cross-linking, the double stranded nucleic acid melted, and the process of base pairing, activation and melting repeated, a sufficient number of cycles, to provide a detectable amount of cross-linked probes. To inhibit background cross-linking, the side chains may provide for duplex formation, where a portion of the side chain binds to a different portion of the side chain or the portion of the probe homologous to the target. Also provided are kits comprising reagents, as well as automatic devices, for carrying out the subject method.
提供了检测核酸序列的方法和组合物。特别是采用了成对探针,其中成对探针定义了目标核酸上基本连续的序列。每对探针都有一条侧链,当探针和样本核酸碱基配对时,两条侧链形成一个茎,茎与探针非共价结合,并能在活化时形成交联。未与互补 DNA 结合时,茎的交联会受到抑制。每种核酸最初都是单链核酸,以便进行碱基配对,从而使探针与同源靶核酸结合。检测混合物被激活以产生交联,双链核酸被融化,碱基配对、激活和融化的过程重复足够多的循环,以提供可检测到的交联探针量。为抑制背景交联,侧链可提供双链形成,其中侧链的一部分与侧链的不同部分或探针与目标同源的部分结合。此外,还提供了包含试剂的试剂盒以及自动装置,用于实施主题方法。