公开(公告)号：US20060014152A1

公开(公告)日：2006-01-19

申请号：US10867994

申请日：2004-06-15

申请人： Martin Mautner

发明人： Martin Mautner

IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/04

CPC分类号： C12Q1/6858 ,  C12Q1/6869 ,  C12Q2563/173 ,  C12Q2527/125 ,  C12Q2535/125

摘要： A method for reducing the efficiency of primer extension by polymerase enzymes when the 3′ end of a primer or growing nucleic acid chain does not hybridize perfectly with the target, for increasing the selectivity of single nucleotide mutation or gene analyses, for suppressing false positive results and for enhancing the fidelity of the amplification of nucleic acid fragments by avoiding the incorporation of mispairs, the method comprising the steps of: (a) obtaining a nucleic acid sample; (b) hybridizing said nucleic acid sample to a primer; (c) subjecting said nucleic acid sample hybridized to a extension reaction by extending a primer with a polymerizing enzyme; and (d) detecting the presence of extension products; wherein the reaction extension mixture medium contains an intercalating agent such as ethidium bromide, dihydroethidium, ethidium homodimer-1, ethidium homodimer-2, acridine, propidium iodide, YOYO®-1 or TOTO®-1. When the intercalating agent is ethidium bromide the concentration is about 1 to 7 μg/ml.

摘要翻译： 当引物或生长核酸链的3'末端与靶完全杂交时，为提高单核苷酸突变或基因分析的选择性，抑制假阳性结果，降低聚合酶引物延伸效率的方法 并且为了通过避免引入错配来增强核酸片段的扩增的保真度，该方法包括以下步骤：（a）获得核酸样品; （b）将所述核酸样品与引物杂交; （c）通过用聚合酶延伸引物对经过延伸反应杂交的所述核酸样品进行处理; 和（d）检测延伸产品的存在; 其中所述反应延伸混合物介质含有插层剂，例如溴化乙锭，二氢乙硫醚，同系二聚体同族二聚体-1，同系二聚体-2，吖啶，碘化丙啶，YOYO或TOTO。 当插入剂是溴化乙锭时，浓度为约1至7mug / ml。