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Help! about transformation E. coli

Help! about transformation E. coli - DNA Techniques

Help! about transformation E. coli - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


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  #1  
Old 01-27-2007, 02:38 PM
Pipette Filler
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Default Help! about transformation E. coli



Hi, all,

I am doing transformation of E. coli with a cDNA vector. I have never done this before. Yesterday I did once but after 12 hour, nothing appeared on the LB plate. So what is the possible reason for this?

Thanks a lot!

sunoom
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  #2  
Old 01-27-2007, 04:15 PM
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Default Re: Help! about transformation E. coli

Hi sunoom
Welcome to the fourm
well there are many ways to transform E.coli cells,
as there is chemical method mostly by use of CaCl2 treatment, E.coli cells are made competent to transform.
and E.coli cells are also transformed by electroporation.
what method you tried for transformation??
there are many reason for not successful transformation
1 the cells may not be fully competent
2 the size of the vector you use for transformation, as big size vector are not easily transformed
3 the cells might be transformed but your selection procedure may be wrong.....
so there are many explainations are there for transformation failure



best regards
aftab
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Old 02-20-2007, 10:58 AM
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Default Re: Help! about transformation E. coli

hi
i agree with aftabac give the total details u have used /followed then possible reason can expect
c u
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