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chloramphenicol storage and amplification


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#1 Sindit

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Posted 02 March 2011 - 11:24 AM

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!

#2 pito

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Posted 02 March 2011 - 11:34 AM

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" ?
If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.

#3 Sindit

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Posted 02 March 2011 - 11:40 AM

Sorry,

I mean thermo shock into a DH5 alpha... but I heard that this E. coli doesn't replicate it...




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" ?



#4 pito

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Posted 02 March 2011 - 11:58 AM

Sorry,

I mean thermo shock into a DH5 alpha... but I heard that this E. coli doesn't replicate it...





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.
If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.




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