How can I analysis the transcript after T7 promoter? - [HELP] (Aug/14/2006 )
Well Doc Martin,
I mean I do not know the start base and the end base.
If I linearize the plasmid, will the transcription stop at the end of the sequence? (There is no palindrome or some stop signal in front)
well, what do you know about the DNA that you have cloned? if you do not know (or at least have a prediction) then your DNA may or may not be in frame and may or may not contain a coding sequence. what I'm saying is, you might not get a transcript at all
what are all the details you know about the DNA fragment you have cloned into the plasmid? anything you know about it, besides the size? have you sequenced it yet?
often times vectors will contain stop codons downstream of the cloning region, and that is where transcription will stop (if there isn't a stop site in your DNA)
I have cloned the segment just after the T7 promoter, and just 10bp lower, the MCS ending.