Boston University/Bacterial Conjugation Results

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

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

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.


BU conjugation2.jpg

Plate 2


BU conjugation3.jpg

Plate 3


BU conjugation4.jpg

Plate 4


BU conjugation5.jpg

Plate 5

Back