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1. 发明授权

US07223536B2 Methods for detecting single nucleotide polymorphisms 失效
标题翻译：检测单核苷酸多态性的方法
公开(公告)号：US07223536B2
公开(公告)日：2007-05-29
申请号：US09778168
申请日：2001-02-07
申请人： David J. Wright , Maria A. Milla , James G. Nadeau , G. Terrance Walker
发明人： David J. Wright , Maria A. Milla , James G. Nadeau , G. Terrance Walker
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/04
CPC分类号： C12Q1/6827 , C12Q2565/1025 , C12Q2535/125 , C12Q2531/119 , C12Q2531/101
摘要： The present invention provides methods for detecting and identifying sequence variations in a nucleic acid sequence of interest using a detector primer. It has been found that the reduced efficiency of primer extension by DNA polymerases when the 3′ end of a primer does not hybridize perfectly with the target can be adapted for use as a means for distinguishing or identifying the nucleotide in the target which is at the site where the diagnostic mismatch between the detector primer and the target occurs. The detector primer hybridizes to the sequence of interest and is extended with polymerase. The efficiency of detector primer extension is detected as an indication of the presence and/or identity of the sequence variation in the target. The inventive methods make use of nucleotide mismatches at or near the 3′ end of the detector primer to discriminate between the nucleotide sequence of interest and a second nucleotide sequence which may occur at that same site in the target. The methods are particularly well suited for detecting and identifying single nucleotide differences between a target sequence of interest (e.g., a mutant allele of a gene) and a second nucleic acid sequence (e.g., a wild type allele for the same gene).
摘要翻译：本发明提供使用检测器引物检测和鉴定感兴趣的核酸序列中的序列变异的方法。已经发现，当引物的3'端没有与靶完全杂交时，通过DNA聚合酶的引物延伸的效率降低可以适用于区分或鉴定靶中的核苷酸的手段发现检测器引物与靶标之间的诊断不匹配的位置。检测引物与感兴趣的序列杂交，并用聚合酶扩增。检测器引物延伸的效率被检测为目标中序列变异的存在和/或身份的指示。本发明的方法利用在检测器引物的3'末端附近或其附近的核苷酸错配来区分目的核苷酸序列和可能发生在靶的相同位点的第二核苷酸序列。所述方法特别适用于检测和鉴定靶标目标序列（例如，基因的突变等位基因）和第二核酸序列（例如，相同基因的野生型等位基因）之间的单核苷酸差异。

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