申请人:University of Utah Research Foundation
公开号:EP2826867A1
公开(公告)日:2015-01-21
The invention provides a method for nucleic acid analysis comprising the steps of
mixing a target nucleic acid with a saturating dsDNA binding dye and an unlabeled probe configured to hybridize to a portion of the target nucleic acid, to form a mixture,
amplifying the target nucleic acid to generate an amplicon,
allowing the unlabeled probe to hybridize to the amplicon to form a probe/amplicon duplex,
generating a melting curve for the probe/amplicon duplex by measuring fluorescence from the dsDNA binding dye as the mixture is heated, and
analyzing the shape of the melting curve.
Also provided are kits for analyzing a target nucleic acid comprising:
oligonucleotide primers configured for amplifying the target nucleic,
a saturating dsDNA binding dye.
a thermostable polymerase, and
acid an unlabeled probe configured to hybridize at least partially to the target nucleic acid.
本发明提供了一种核酸分析方法,包括以下步骤
将目标核酸与饱和 dsDNA 结合染料和配置为与目标核酸部分杂交的未标记探针混合,形成混合物、
扩增目标核酸以产生扩增子、
让未标记探针与扩增子杂交,形成探针/扩增子双链体、
通过测量混合物加热时 dsDNA 结合染料发出的荧光,生成探针/扩增子双链体的熔化曲线,以及
分析熔化曲线的形状。
还提供了用于分析目标核酸的试剂盒,包括
用于扩增目标核酸的寡核苷酸引物、
饱和 dsDNA 结合染料。
一种恒温聚合酶,以及
酸 一个未标记探针,配置为至少部分与目标核酸杂交。