amp plates containing xgal/iptg with expression vector
Posted 03 October 2011 - 01:55 PM
I'm planning on ligating my gene into an expression vecot (Pet32a) and have to plate on amp plates--i made some up a while ago containing xgal/iptg so just wondering if i can use these...i know i wont get blue colonies..but having these substances added won't make any difference to the end result right?
Posted 03 October 2011 - 10:10 PM
Posted 04 October 2011 - 05:21 AM
Best of Luck.
Posted 04 October 2011 - 09:54 PM
Posted 04 October 2011 - 11:30 PM
Thanks for both your explantations...allynspear: i read your post and was like ah ok, so i can continue using those plates..UNTIL I read your last sentace ...BL21 is what I'm using haha...but it's all good..I just made a bunch of new plates with amp/chloramphenicol...didn't end up getting any colonies using those other plates anyway so thought may as well make a whole new bunch..
protolder: no need to be sorry at all ..your answer was helpful as well
Posted 05 October 2011 - 05:46 AM
Biology_06er, I am wondering why you are transforming a ligation into BL21 cells, though. BL21 are an expression cell line that you normally would transform into once you have verifyied your clone. They don't usually give high quality mini/maxi prep DNA, and they tend to be more expensive than cloning-grade competent cells. It's not that you CAN'T do it, just wondering if there was some specific reason.
Best of Luck.
Posted 05 October 2011 - 12:56 PM
I first transformed into omnimax cells and then miniprepped (and now currently) trying to cut my gene out for my pbs vector and into an expression vector...it didn't work the first time (hence my other post regarding transforming straight into BL21 from pcr product)..I'm trying again today the gel purification techniquew as that is much better