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problem with expression vector cultivation

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

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problem with expression vector cultivation

Postby genetherapy » Sat Mar 03, 2012 11:26 am

I use pfastBAC1 expression vector from invitrogen. I cultivate 15 mikroliter original vector to 200 ml ampicilin tb which the vector have the resistance gene. I wanted to stock the vector but ı have problems with these copy vectors. I do my ligations with this copy vectors that ı cultivated from original but none of my ligations performed. So I decided to make an experiment if the problem is with my ligase or my cultivated plasmids,I spread cultivated vector to ampicilin plate, I also spread the digested cultivated vector to ampicilin plate to test if ligase works or not. I looked my plates today but where were no colonies so this means the problem is with my cultivated vector BUT I REALLY DO NOT UNDERSTAND WHY I CAN NOT USE CULTIVATED PLASMIDS. ISN'T IT POSSIBLE TO CULTIVATE VECTOR FROM ORIGINAL (COMMERCIAL) VECTORS? DO ANYONE HAVE AN OPINION ABOUT THESE OR HAVE SOME EXPERIENCE?
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Postby JackBean » Fri May 18, 2012 8:52 am

by cultivating vector you mean bacteria harboring that vector, right?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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