涂7体外转录翻译汇编.ppt

In vitro transcription and In vitro translation In vitro Transcription In vitro Transcription Plasmid with T3, T7 or SP6 promoters Linearization of plasmid DNA by restriction enzyme In vitro transcription: Plasmid buffer NTP labeled UTP RNA polymerase DNAse digestion, Phenol/Chloroform extraction and RNA precipitataion Advantages of RNA probes: RNA-RNA hybrids are very stable Tissue can be digested with RNase (dsRNA is not digested) after the hybridization reducing the background Higher specific activity compared to oligonucleotides 蛋白质功能研究思路 DNA水平缺失基因,上调下调基因的表达水平对生命过程的影响