公开(公告)号：EP0863998A1

公开(公告)日：1998-09-16

申请号：EP96937086.0

申请日：1996-11-01

申请人： NAXCOR

发明人： WOOD, Michael ,  VAN ATTA, Reuel ,  ALBAGLI, David

IPC分类号： C12N15 ,  C12Q1

CPC分类号： C12Q1/6827 ,  Y10S435/81 ,  C12Q2565/131 ,  C12Q2523/101 ,  C12Q2523/313 ,  C12Q2565/125

摘要： Double-stranded conformational polymorphism analysis is performed by combining a probe comprising a cross-linking agent and optionally a label with a sample having a target sequence, which may be complementary or have one or a few mismatches with respect to the probe sequence. After sufficient time for hybridization under mild or lesser stringency conditions, hybridized pairs are irradiated to induce cross-link formation by the cross-linking agent. The sample is then analyzed by denaturing gel electrophoresis where the rate of migration depends upon the degree of complementarity between the probe and the target. For corroboration, in a second experiment, the probe may be combined with the sample under high stringency conditions, where it is found that the formation of cross-linked probe/target is substantially lower for pairs having mismatches than for fully matched pairs. After cross-linking, the sample may be separated by gel electrophoresis, and the amount of cross-linked nucleic acid determined.

公开(公告)号：EP0796346A1

公开(公告)日：1997-09-24

申请号：EP95944246.0

申请日：1995-12-22

申请人： NAXCOR

发明人： ALBAGLI, David ,  VANATTA, Reuel ,  WOOD, Michael

IPC分类号： G01N33 ,  B01L7 ,  C07H21 ,  C12M1 ,  C12N15 ,  C12P19 ,  C12Q1

CPC分类号： C12Q1/6816 ,  B01L7/52 ,  B01L2300/1883 ,  Y10S435/81 ,  C12Q2525/301 ,  C12Q2523/101 ,  C12Q2537/119 ,  C12Q2523/313

摘要： 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.