frustrating maxiprep problem - (Jun/25/2008 )
I am working on a dozen of maxiprep from palsmids. But some strange things keep happening. Most of them grew very well on Amp+ LB plates after transfromation. But some of them don't grow in 3ml Amp LB media. Among those who grow in LB media, some of them no plasmid after miniprep. What's happening?
How long do u keep your plates for? If you are keeping it for more than 16-18 hours, there are chances the colonies you are picking are satellite colonies.
Metz
A couple of us have been talking about this problem in another post:
http://www.protocol-online.org/forums/inde...showtopic=37242
I'm looking into it. Can you give me the details of your vector and insert?
Cheers, Rob
hallo, if you try to clone gene , product of is toxic for cell it is possinble the cell get it off. or ATP is degradated. Apiciline plate and media suppose to be used to one week
good luck
Those plates were kept for 2 days. I understand that satellite colony can grow on the plate, but how can a satellite colony (if I unluckily pick them from the plates) grow in fresh Amp LB media?
How long do u keep your plates for? If you are keeping it for more than 16-18 hours, there are chances the colonies you are picking are satellite colonies.
Metz