公开(公告)号：US20060094009A1

公开(公告)日：2006-05-04

申请号：US10519153

申请日：2003-06-25

申请人： Patrick Vaughan ,  Thomas McCarthy

发明人： Patrick Vaughan ,  Thomas McCarthy

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/683 ,  C12Q1/6858 ,  C12Q2531/113 ,  C12Q2525/131 ,  C12Q2525/101 ,  C12Q2521/301

摘要： A method for characterising nucleic acid molecules comprises the steps of i) introducing a modified base which is a substrate for a DNA endonuclease, for example Endonuclease V from E. coli, into a target DNA molecule; and ii) reacting the nucleic acid containing the modified base with said DNA endonuclease such that the nucleic acid is cleaved to generate an upstream fragment condition said modified base and bearing a 3′ hydroxyl group. The upstream fragment generated in step ii) can be used as a primer for a subsequent extension reaction. The modified base as such targets cleavage of the nucleic acid strand because it directs the endonuclease to cut the nucleic acid strand on which it resides at a site predominantly one nucleotide removed therefrom, so that the modified base remains in the nucleci acid as th upstream fragment. The method can be used for detecting polymorphisms and mutations and for nucleic acid indentification and profiling. The method enables one to detect polymorphisms in any sequence context and such detection is not dependent on surrounding nucleotide sequence.

摘要翻译： 用于表征核酸分子的方法包括以下步骤：i）将作为DNA内切核酸酶的底物的修饰的碱（例如来自大肠杆菌的核酸内切酶V）引入靶DNA分子中; 和ii）使含有修饰碱基的核酸与所述DNA内切核酸酶反应，使得所述核酸被切割以产生上游片段条件，所述修饰碱基并带有3'羟基。 在步骤ii）中产生的上游片段可以用作随后的延伸反应的引物。 修饰的碱基靶向核酸链的切割，因为其指导内切核酸酶切割其所在的位于主要从其去除的一个核苷酸的位点处的核酸链，使得修饰的碱基作为上游片段保留在核酸中 。 该方法可用于检测多态性和突变以及核酸识别和分析。 该方法使得能够检测任何序列上下文中的多态性，并且这种检测不依赖于周围的核苷酸序列。

公开(公告)号：USD327857S

公开(公告)日：1992-07-14

申请号：US541363

申请日：1990-06-21

申请人： Patrick Vaughan

设计人： Patrick Vaughan