A PCR-based strategy for cloning short hairpin sequences: PCR shagging

The overall approach is to use an RNA polymerase III promoter to drive
expression of encoded short hairpin RNA (shRNA). For this purpose we use the U6 snRNA promoter and maintain the transcript initiating G nucleotide of the U6snRNA transcript. There by, hairpin sequences will start with a G. Termination is mediated by a run of Ts at the end of the hairpin.

Protocol (PDF file) written by Patrick J. Paddison, Watson School of Biological Sciences, Cold Spring Harbor Lab.

Last update 30-Jan-2005, Rating n/a of 0 votes.