Extraction and purification of plasmid DNA Agarose gel electrophoresis Restriction enzyme digestion Recovery and purification of DNA fragment on agarose gel Phenol/chloroform extraction and alcohol precipitation DNA ligation Preparation and transformation of competent E. coli Identification of bacterial colonies that contain recombinant plasmids Polymerase chain reaction (PCR)
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Bacteria culture
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Extraction and purification of plasmid DNA
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Gene cloning
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Transformation
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Identification of bacterial colonies that contain recombinant plasmids
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PCR
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SDS-PAGE
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Western blotting
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