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1. 发明申请

WO2016164363A8 METHODS FOR DETERMINING BASE LOCATIONS IN A POLYNUCLEOTIDE 审中-公开
标题翻译：用于确定多核苷酸中碱基位置的方法
公开(公告)号：WO2016164363A8
公开(公告)日：2016-12-01
申请号：PCT/US2016026047
申请日：2016-04-05
申请人： UNIV CALIFORNIA
发明人： JAIN MITEN , OLSEN HUGH EDWARD , AKESON MARK A
IPC分类号： C12M1/34 , C12N11/00 , C12N15/10
CPC分类号： C12Q1/6869 , C12Q1/6827 , C12Q2521/531 , C12Q2525/119 , C12Q2545/101 , C12Q2563/116 , C12Q2565/631 , C12Q2537/164
摘要： Disclosed are methods for polynucleotide sequencing that detect the location of selected nucleobases with greater precision. The methods can be used to determine the location and nature of modified bases in a polynucleotide, that is, non-canonical bases, or to improve accuracy of sequencing of "problem" regions of DNA sequencing such as homopolymers, GC rich areas, etc. The sequencing method exemplified is nanopore sequencing. Nanopore sequencing is used to generate a unique signal at a point in a polynucleotide sequence where an abasic site (AP site, or apurinic or apyrimidinic site) exists. As part of the method, an abasic site is specifically created enzymatically using a DNA glycosylase that recognizes a pre-determined nucleobase species and cleaves the N-glycosidic bond to release only that base, leaving an AP site in its place.
摘要翻译：公开了用于多核苷酸测序的方法，其以更高的精度检测所选核碱基的位置。这些方法可用于确定多核苷酸（即非规范碱基）中修饰碱基的位置和性质，或提高DNA测序“问题”区域测序的准确性，如均聚物，GC富集区等。示例的测序方法是纳米孔测序。纳米孔测序用于在存在脱碱基位点（AP位点或无嘌呤或脱嘧啶位点）的多核苷酸序列中的某一点产生唯一信号。作为该方法的一部分，使用识别预定核碱基物种的DNA糖基化酶酶切地形成脱碱基位点，并切割N-糖苷键以仅释放该碱基，从而将AP位点置于其位置。

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