公开(公告)号：US07459311B2

公开(公告)日：2008-12-02

申请号：US10363231

申请日：2001-09-07

申请人： Pål Nyren ,  Mostafa Ronaghi ,  Annika Tallsjö

发明人： Pål Nyren ,  Mostafa Ronaghi ,  Annika Tallsjö

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6869 ,  C12Q2565/301

摘要： The present invention provides a method of identifying a base at a target position in a sample nucleic acid sequence, said method comprising: subjecting a primer hybridised to said sample nucleic acid immediately adjacent to the target position, to a polymerase primer extension reaction in the presence of a nucleotide, whereby the nucleotide will only become incorporated if it is complementary to the base in the target position, and determining whether or not said nucleotide is incorporated by detecting whether Ppi is released, the identity of the target base being determined from the identity of any nucleotide incorporated, wherein, where said nucleotide comprises an adenine base, an α-thio triphosphate analogue of said nucleotide is used, and the Rp isomer of said analogue and/or the degradation products of said analogue are eliminated from the polymerase reaction step.

摘要翻译： 本发明提供了鉴定样品核酸序列中目标位置的碱基的方法，所述方法包括：使与所述目标位置相邻的所述样品核酸杂交的引物在存在的情况下进行聚合酶引物延伸反应 的核苷酸，其核苷酸仅在与目标位置的碱基互补时才被引入，并且通过检测Ppi是否被释放来确定所述核苷酸是否被并入，靶基因的身份是根据身份确定的 其中，当所述核苷酸包含腺嘌呤碱基时，使用所述核苷酸的α-硫代三磷酸类似物，并且所述类似物的Rp异构体和/或所述类似物的降解产物从聚合酶反应步骤中消除 。