Journal of the Association for Laboratory

摘要: Current applications of Q-PCR for the quantitative analysis of mRNA expression involve the following: Total RNA isolation, first strand cDNA synthesis, Q-PCR amplification, measurement of fluorescence accumulation, data extraction, and data analysis. Each of these steps can be both laborintensive and time-consuming. In particular, the extraction of total RNA from a large number of samples and the subsequent synthesis of first strand cDNA typically involve numerous pipetting and centrifugation steps which limits their application to automated high-throughput systems. Here, we describe the use of a robotic workstation, integrated with a bank of thermocyclers and a capillary electrophoresis unit, for the coupled extraction of total RNA, first strand cDNA synthesis, and quality control monitoring. As a result, manual labor, total procedure time, error rate, and ‘‘drop-outs’’are reduced to a minimum. We have further increased the high-throughput nature of this system by pairing these robotically assisted pipetting steps with an automated data extraction and