chloramphenicol storage and amplification - (Mar/02/2011 )
Hello,
I have a sample of pKD3 plasmid (chloramphenicol resistance) and I need to put it into some bacteria to keep and reproduce it. I realized that it doesn't work with regular transformation so... does anyone have a protocol, procedure, reference or even tips for that?
It's so simple but I couldn't found anything on the web.
Thanks!
-Sindit-
Sindit on Wed Mar 2 19:24:31 2011 said:
Hello,
I have a sample of pKD3 plasmid (chloramphenicol resistance) and I need to put it into some bacteria to keep and reproduce it. I realized that it doesn't work with regular transformation so... does anyone have a protocol, procedure, reference or even tips for that?
It's so simple but I couldn't found anything on the web.
Thanks!
What do you mean with "regular transformation" ?
-pito-
Sorry,
I mean thermo shock into a DH5 alpha... but I heard that this E. coli doesn't replicate it...
pito on Wed Mar 2 19:34:46 2011 said:
Sindit on Wed Mar 2 19:24:31 2011 said:
Hello,
I have a sample of pKD3 plasmid (chloramphenicol resistance) and I need to put it into some bacteria to keep and reproduce it. I realized that it doesn't work with regular transformation so... does anyone have a protocol, procedure, reference or even tips for that?
It's so simple but I couldn't found anything on the web.
Thanks!
What do you mean with "regular transformation" ?
-Sindit-
Sindit on Wed Mar 2 19:40:33 2011 said:
Sorry,
I mean thermo shock into a DH5 alpha... but I heard that this E. coli doesn't replicate it...
pito on Wed Mar 2 19:34:46 2011 said:
Sindit on Wed Mar 2 19:24:31 2011 said:
Hello,
I have a sample of pKD3 plasmid (chloramphenicol resistance) and I need to put it into some bacteria to keep and reproduce it. I realized that it doesn't work with regular transformation so... does anyone have a protocol, procedure, reference or even tips for that?
It's so simple but I couldn't found anything on the web.
Thanks!
What do you mean with "regular transformation" ?
The pKD3 plasmid needs to have lambda pir protein...do you this have DH5pir or DH5?
try electroporation, this might to the tric.
-pito-