Problem with shRNA sequencing! Help. - (Feb/03/2007 )
I'm trying for last weeks to sequence my vector with inserted shRNA. The vector is LMP retroviral vector. I confirmed insertion by many digestions, but when comming to sequencing it looks realy bad. I need to check the sequence of the linkers, does anyone has some good tips, and how much DNA I should use?
You have to check with the sequencing center about sequencing shRNA vectors. Our center advices to give them 800ng of DNA and 10pmol of primer and they add DMSO and perform a special PCR protocol for shRNA vectors. It works perfectly fine.
Thank you. Is it possible to get that PCR protocol from you?
its not possible, as the core have established the protocol.