Belin D
Department of Pathology, University of Geneva Medical School, CMU, Switzerland.
Mol Biotechnol. 1997 Apr;7(2):153-63. doi: 10.1007/BF02761751.
The monomeric bacteriophage RNA polymerases allow the synthesis of virtually any RNA molecule in unlimited quantity. In this protocol, we describe the preparation of plasmid and PCR-derived templates. A basic transcription protocol is provided with several optional modifications. The use of RNA probes in Northern blot hybridization and in RNase protection assays is described. The relative advantages and pitfalls of these two methods to quantitatively detect mRNA targets are discussed.
单体噬菌体RNA聚合酶能够无限量地合成几乎任何RNA分子。在本实验方案中,我们描述了质粒和PCR衍生模板的制备方法。还提供了一个基本的转录实验方案以及几种可选的改进方法。阐述了RNA探针在Northern印迹杂交和RNA酶保护分析中的应用。讨论了这两种定量检测mRNA靶标的方法的相对优点和缺陷。