From 2007.igem.org
Filter Conjugation Protocol
- Filter conjugation to transfer the plasmid pMS291(lacZ) from E.coli SM10 to S. oneidensis AS-92.
- The plasmid pMS291 contained a kanamycin resistance gene. S. oneidensis has a native gentamicin resistance.
- The plasmid pMS291 is first transformed into E.coli SM10 and then the conjugation protocol is run to encourage conjugal transfer of the plasmid to S. oneidensis AS-92. A double-antibiotic selection method is used to select for successful S. onedensis transformants which have a resistance for both kanamycin and gentamicin.
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