From 2007.igem.org

Read more about how conjugation works

Our 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.

Plate 1: 7 separate conjugation mixes were plated. The white is the filter upon which the bacteria conjugate. S. oneidensis is red.

Plate 2

Plate 3

Plate 4

Plate 5

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