公开(公告)号：US20050053950A1

公开(公告)日：2005-03-10

申请号：US10658602

申请日：2003-09-08

申请人： Enrique Zudaire Ubani ,  Frank Cuttitta

发明人： Enrique Zudaire Ubani ,  Frank Cuttitta

IPC分类号： B01L7/00 ,  C12P19/34 ,  C12Q1/68

CPC分类号： C12Q1/6851 ,  B01L7/52 ,  C12Q1/6858 ,  C12Q1/686 ,  C12Q2561/113 ,  C12Q2537/143 ,  C12Q2527/107

摘要： The present invention provides a new protocol for quantifying multiplex real-time polymerase chain reaction (PCR). In particular, the present invention provides methods of quantifying multiple PCR products or amplicons in a single real-time PCR reaction based on the different melting temperatures (Tm) of each amplicon and the emission changes of double stranded DNA dyes such as SYBR Green I when amplicons are in duplex or in separation. For a specific amplicon with a Tm, the emission difference between the emission reading taken at a temperature below the Tm and the emission reading taken at a temperature above the Tm corresponds to the emission value of the amplicon in duplex. Accordingly, the emission difference of each amplicon in a single PCR reaction can be used to quantify each amplicon. The present invention further provides computer programs or computer products which perform the methods described herein.

摘要翻译： 本发明提供了一种用于量化多重实时聚合酶链反应（PCR）的新方案。 特别地，本发明提供了基于每个扩增子的不同解链温度（Tm）和双链DNA染料如SYBR Green I的发射变化在单次实时PCR反应中定量多个PCR产物或扩增子的方法， 扩增子是双链体或分离的。 对于具有Tm的特异性扩增子，在低于Tm的温度下获取的发射读数与在高于Tm的温度下获得的发射读数之间的发射差异对应于双链体扩增子的发射值。 因此，单个PCR反应中每个扩增子的发射差异可用于量化每个扩增子。 本发明还提供了执行本文描述的方法的计算机程序或计算机产品。