Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Cloning

emty vector cloning - (Jan/13/2012 )

What is the proper procedure to confirm successful cloning of a vector that is without insert? Do I cut with just one restriction enzyme? Thanks for helping.


Your question was not carrining enough details to solve your problem.

Let me try to answer.

Check cloning sucess is by Restriction digestion, but i didnot understand what is without insert, if it is just plasmid. Then you have to just transform the plasmid into bacterial and you are done.


Thanks, this is exactly what I meant. I have just the plsmid, which I needed more of, so I transformed E. coli, did minipreps, and was wandering if I should somehow check the plasmid.


Yes, I would check the plasmid by restriction digest. However, I'd suggest to use a multiple cutting enzyme (at least 2x), which gives you a better idea of the identity of your plasmid. I really like BanII, which cuts GRGCY^C, and thus most vectors at least once.