Parameters involved in the conversion of real-time PCR assays from the ABI prism 7700 to the Cepheid SmartCycler® II

被引：7

作者：

Bentley, HA ^{[1
]}

Belloni, DR ^{[1
]}

Tsongalis, GJ ^{[1
]}

机构：

[1] Dartmouth Hitchcock Med Ctr, Dartmouth Med Sch, Dept Pathol, Lebanon, NH 03766 USA

来源：

CLINICAL BIOCHEMISTRY | 2005年 / 38卷 / 02期

关键词：

D O I：

10.1016/j.clinbiochem.2004.10.009

中图分类号：

R446 [实验室诊断]; R-33 [实验医学、医学实验];

学科分类号：

1001 ;

摘要：

Objective: We examined several critical parameters that must be optimized when converting between the ABI Prism 7700 real-time PCR platform and the Cepheid SmartCycler((R)) II while using the same primer and probe sequences. Design and methods: A lyophilized master mix, MgCl2 concentration, PCR cycling conditions, and ramp times were evaluated. Results: Optimization of each parameter, including use of the OmniMix(TM) HS-lyophilized beads, 6 mM MgCl2 concentration, changes in PCR cycling parameters, and increased ramp time were necessary to convert this real time PCR assay to a new platform. Conclusion: We conclude that careful consideration of several analytical parameters can result in a smooth transition of assays between real time PCR platforms. (C) 2004 The Canadian Society of Clinical Chemists. All rights reserved.

引用

页码：183 / 186

页数：4

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