Protocol Online logo
Top : Forum Archives: : Molecular Biology

sequencing vector DNA - (Feb/21/2008 )

hey, my question is concerning when would be a good time to sequence the gene of interest on the vector. so someone has already cloned my gene of interest and ligated it into the ptarget vector and verified it was the gene of interest through sequencing. he has transformed the vector into e. coli and purified vector using a maxi prep; however, that was several months ago. i recently re-transformed some more e. coli using that maxi prep'd vector (because i was running low on the gene-ptarget vector) and isolated more vector again. do i have to sequence again?


I probably wouldn't bother sequencing it again, but if you're concerned that its not the right thing then you might do a restriction digest to make certain that it at least looks right. If you start to have problems with it then sequencing might be warranted.


Do you have his lab books? Can you see his data?

If you can see the sequencing trace, digest the DNA to show it is the right thing, just incase there is a mistake in the labelling.

If you can't see his data, sequence it again. The painfull leason that you will learn is trust nobody, not even yourself. Anything that can go wrong will go wrong, especially when there is an audience to display itself too.




As pernseblue suggested, look at the sequence yourself, if its not there, then sequence it.