Stoeckli Esther T
CSH Protoc. 2006 Sep 1;2006(4):pdb.prot4503. doi: 10.1101/pdb.prot4503.
INTRODUCTIONThis protocol describes the production of double-stranded RNA (dsRNA) from fragments of cDNAs of candidate genes. The cDNA fragments must be cloned in plasmids with a flanking SP6 and T7 promoter (e.g., pSP72 or pCRII). The plasmid is linearized and sense and antisense RNAs are produced separately by in vitro transcription. After purification, the RNA strands are annealed to yield a dsRNA molecule suitable for RNAi in avian embryos.
引言
本方案描述了从候选基因的cDNA片段制备双链RNA（dsRNA）的方法。cDNA片段必须克隆到带有侧翼SP6和T7启动子的质粒中（例如，pSP72或pCRII）。将质粒线性化，通过体外转录分别产生正义和反义RNA。纯化后，将RNA链退火以产生适用于禽类胚胎RNA干扰的dsRNA分子。
