Login

Join for Free!
25820 members


Shipping of transformed E.coli

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

Moderator: BioTeam

Shipping of transformed E.coli

Postby Leia » Thu Apr 02, 2009 7:59 am

Hi,
I just recieved some E.coli with transformed pet-plasmids. They were just sent in normal LB-medium and they have been in transit for about 6 days at room temp. Is that a good way to send them? Don't you risk loosing the plasmid when they can just grow for so many days - especially when they are ampicillin resistant and the ampicillin is broken down?

I hope someone can answer this! Thanks!

/Signe
Leia
Garter
Garter
 
Posts: 1
Joined: Thu Apr 02, 2009 7:53 am

Postby MrMistery » Thu Apr 02, 2009 1:49 pm

well did the LB include amp? of course it's a really bad idea if there's no amp in the medium, because there is no pressure for the cells to keep the plasmid.
but i think you can just plate them on an LB+amp plate, pick colonies, do liquid cultures of say 10 colonies, miniprep the cultures and sequence the plasmids. at least some should have the right plasmid, even if there was a contaminant. Just make sure you have isolated individual colonies when you first plate them
"I have no intention of stopping anytime soon. I want to understand the universe and answer the big questions, that is what keeps me going" - Stephen Hawking
User avatar
MrMistery
Inland Taipan
Inland Taipan
 
Posts: 6740
Joined: Thu Mar 03, 2005 10:18 pm
Location: Romania(small and unimportant country)


Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 0 guests