评估 LightCycler 1536 仪器用于高通量实时定量 PCR。

Evaluation of the LightCycler 1536 Instrument for high-throughput quantitative real-time PCR.

机构信息

Group Cardiovascular Genetics, Department of Vertebrate Genomics, Max Planck Institute for Molecular Genetics, Berlin, Germany.

出版信息

Methods. 2010 Apr;50(4):S19-22. doi: 10.1016/j.ymeth.2010.01.007.

DOI:10.1016/j.ymeth.2010.01.007

PMID:20215016

Abstract

Quantitative real-time PCR (qPCR) is a frequently used, sensitive and accurate method to study gene expression profiles. However, its throughput was so far limited for routine laboratories to 384 reactions per run based on the limitations of the available instruments. Recently, the LightCycler 1536 Instrument was launched providing a high-throughput solution for qPCR with the analysis of 1536 reactions in approximately 45 min. We assessed the accuracy and sensitivity of this novel technology for the analysis of gene expression profiles in combination with the Innovadyne Nanodrop Express pipetting robot. We compared expression profiles obtained for 42 genes in 71 samples between the Universal ProbeLibrary and the LightCycler 1536 Instrument and SYBR Green I and the ABI PRISM 7900HT system. We found that the results were highly reproducible between both systems. Beside the higher throughput, the advantage of the LightCycler 1536 Instrument was the reduced consumption of reagents and sample material.

摘要

实时荧光定量 PCR（qPCR）是一种常用的、敏感且准确的方法，用于研究基因表达谱。然而，由于现有仪器的限制，其通量一直限制在每个运行 384 个反应，这对常规实验室来说是不够的。最近，推出了 LightCycler 1536 仪器，为 qPCR 提供了高通量解决方案，大约 45 分钟内可分析 1536 个反应。我们评估了这项新技术与 Innovadyne Nanodrop Express 移液机器人结合用于分析基因表达谱的准确性和灵敏度。我们比较了在 71 个样本中 42 个基因的表达谱，使用了通用探针库和 LightCycler 1536 仪器以及 SYBR Green I 和 ABI PRISM 7900HT 系统。我们发现两个系统之间的结果具有高度的可重复性。除了通量更高之外，LightCycler 1536 仪器的优势还在于减少了试剂和样品材料的消耗。