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Co-transformation


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#1 ahmadfathi

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Posted 09 March 2013 - 02:43 AM

hello, i already done the transformation of my insert into new vector in dh5a host and extracted into a plasmid (CYP3A4-pcWori-dh5a plasmid). then my lecturer said that i need to perform a co-transformation in oxy-reductase vector (pCYAC-OXR) before express the protein, so can i just do the co-transformation of CYP3A4-pcWori-dh5a (Amp resistant) and pCYAC-OXR ( chloramphenicol resistant) plasmid like doing standard transformation? or there have different methods.
now, i just do the standard transformation by put in 25ng of CYP3A4-pcWori-dh5a+ 25ng pCYAC-OXR into dh5a competent cell and streak on chloramphenicol 100ug/ul plate..the result is negative.

#2 pcrman

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Posted 09 March 2013 - 09:54 AM

Never heard of cotransformation. What is its purpose. You do transformation in order to obtain pure colonies which only carry one type of plasmid

#3 ahmadfathi

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Posted 26 March 2013 - 10:45 PM

thank you for your opinion, the purpose of co-transformation is to insert another plasmid into a host which is will help my vector-insert to be expressed well. Right now, i already got it by performing standard transformation with combination of two plasmid in less than 10% of competent cell volume..thank you very much




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