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-	[[Boston University/Filter Conjugation Protocol | Filter Conjugation Protocol]]	+	===Filter conjugation to transfer the plasmid pMS291(lacZ) from E.coli SM10 to S. oneidensis AS-92===
-	FIlter conjugation to transfer the plasmid () from E.coli SM10 to Shewanella AS-92	+	:The plasmid pMS291 contained a kanamycin resistance gene.  S. oneidensis has a native gentamicin resistance.
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		+	: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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		+	[[Boston University/Conjugation | Read more about how conjugation works]]
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		+	[[Boston University/Filter Conjugation Protocol | Our Protocol]]
	[[Image:BU_conjugation1.jpg]]		[[Image:BU_conjugation1.jpg]]
		+
		+	Plate 1: 7 separate conjugation mixes were plated. The white is the filter upon which the bacteria conjugate.  S. oneidensis is red.
		+
		+
	[[Image:BU_conjugation2.jpg]]		[[Image:BU_conjugation2.jpg]]
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		+	Plate 2
		+
		+
	[[Image:BU_conjugation3.jpg]]		[[Image:BU_conjugation3.jpg]]
		+
		+	Plate 3
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		+
	[[Image:BU_conjugation4.jpg]]		[[Image:BU_conjugation4.jpg]]
		+
		+	Plate 4
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		+
	[[Image:BU_conjugation5.jpg]]		[[Image:BU_conjugation5.jpg]]
		+
		+	Plate 5
	[[Boston University | Back]]		[[Boston University | Back]]

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Latest revision as of 16:41, 16 July 2007

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.

Read more about how conjugation works

Our Protocol

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

Back