amp v kan/neo resistance query - (Sep/11/2006 )

Hi,
I've been working with a vector that i didn't have a sequence with, or an adequate map. i've been having problems with getting my things into the right plasmid (i recently posted about having to cancel mice, etc had a nervouse break down and what-not). anyway, i just found out today (!!!!) that my vector doesn't have amp resitance, but is actually kan/neo resistant.
(WOULD HAVE LIKED TO KNOW THAT 3 MONTHS AGO!!!)

anyway, i was wondering, i have grown these plasmids up on an amp plate ... what should have happened?
i would have thought, nothing would grow. yet, i have a lawn.
i'm in the process of checking that the vector in my bacteria is the vector i want... (if it's contamination, i'll die ...again).

V

Take a close look at the data sheet for the strain of bacteria you are using. This happened to me before I realized the strain itself was amp resistant. Also, you can try plating just the bacteria (no plasmid) to see if it grows.

I use JM109s, not amp resistant.
i've got contamination (2 plasmids where there was supposed to only 1) yay for me.

V

Ahhh... contamination. I feel your pain.. especially when my miniprep picked up my fellow labmate's construct. Umm.. can you please NOT contaminate the entire kit next time???