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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.

-salvia-

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.

-Biouday-

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.

-salvia-

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.

-Rsm-