Walters Dana M, Rouvière Pierre E
Central Research and Development, E. I. DuPont de Nemours Co., Wilmington, DE, USA.
Methods Mol Biol. 2006;317:85-97. doi: 10.1385/1-59259-968-0:085.
A high-throughput approach to prokaryotic differential display has been developed. A large number of reverse transcription polymerase chain reactions (RT-PCR) are performed on total RNA isolated from induced and control bacterial cultures. Each RT-PCR reaction uses a single oligonucleotide primer and constitutes an independent sampling of the mRNA population. The large number of reactions performed allows the repeated sampling of the targeted polycistronic mRNA, which is clearly identified among possible false positives.
已开发出一种用于原核生物差异显示的高通量方法。对从诱导培养和对照培养的细菌中分离出的总RNA进行大量逆转录聚合酶链反应(RT-PCR)。每个RT-PCR反应使用单个寡核苷酸引物,构成对mRNA群体的独立采样。所进行的大量反应允许对靶向的多顺反子mRNA进行重复采样,这在可能的假阳性中能被清晰识别。