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Electroporation to generate knockout

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#1 tofuj



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Posted 22 July 2015 - 05:55 PM

Hi all! I am looking to do electroporation to generate a knockout in a particular bacterial species. Briefly, what I have done is created a PCR product composed of 5'flanking region of my gene of interest - antibiotic casette - 3' flanking region of my gene of interest, which I will electroporate into the bacteria and hopefully get homologous recombination to replace my gene of interest with the antibiotic resistant gene. 


My question is if it will be alright just to electroporate my PCR product (~1.4kb) without first cloning it into a vector? 

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