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emty vector cloning

cloning empty vector

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3 replies to this topic

#1 salvia

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Posted 13 January 2012 - 01:41 AM

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.

#2 Biouday

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Posted 13 January 2012 - 02:56 AM

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.

#3 salvia

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Posted 13 January 2012 - 03:03 AM

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.

#4 Rsm

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Posted 13 January 2012 - 03:49 AM

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.
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