发明申请
- 专利标题： Protocol and software for multiplex real-time PCR quantification based on the different melting temperatures of amplicons
- 专利标题（中）： 基于扩增子的不同融合温度的多重实时PCR定量方法和软件
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申请号： US10658602申请日： 2003-09-08
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公开(公告)号： US20050053950A1公开(公告)日： 2005-03-10
- 发明人: Enrique Zudaire Ubani , Frank Cuttitta
- 申请人： Enrique Zudaire Ubani , Frank Cuttitta
- 主分类号： B01L7/00
- IPC分类号： B01L7/00 ; C12P19/34 ; C12Q1/68
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.