Boston University/Electroporation Results

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==Introduction to Electroporation and Testing==
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:During the weekend of 7/14/07, the BU iGEM team ran a test of Electroporation of plasmids pBAD18s, pBAD30, and pjQ200 into wildtype S. oneidensis. Plasmids pBAD18s and pBAD30 were recommended to us by Professor Gardner as possible vectors for mutated Shewy genes. pjQ200 was the plasmid our team had previously selected as a prime candidate for serving as a vector. In order to check (and in the case of pjQ200, double-check) that these plasmids are capable of direct transformation into Shewy, tests of Zymo transformation and electroporation were conducted. For both types of transformation, E. coli was used as a control to verify that our transformation procedures were conducted properly. The results are posted below in a series of images and a short analysis. Lastly, gentamycin plates were used to select for pjq200 transformants, while ampicillin plates were used to select for pBAD18s and pBAD30 transformants.
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==Results of Electroporation==
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{| style="width:700pt;" border="1"
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|
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::::'''E. Coli Transformants'''
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:The image immediately to the right of this text shows the results from our transformation of E.coli with plasmids pBAD18s, pBAD30, and pjQ200 via Zymo transformation and electroporation.
 +
 +
:The contents of the plates are as follows(column indicates transformation type):
 +
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{| align="center" border="1"
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|'''Zymo(poured)'''||'''Zymo(streaked)'''||'''Electroporation'''
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|-
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|pBAD18s|||pBAD18s|||pBAD18s
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|-
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|pBAD30|||pBAD30|||pBAD30
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|-
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|pjQ200|||pjQ200|||pjQ200
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|}
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:The "poured" and "streaked" designations refer to how the transformed E. coli were put onto the plate. When transformed E. coli were streaked onto the plate, 5 microliters were used. When transformed E. coli were poured onto the plate, 100 microliters were used. Electroporated E. coli were all poured onto plates.
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|
[[Image:BU_ecoli_zymo_electro.jpg]]
[[Image:BU_ecoli_zymo_electro.jpg]]
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|-
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 +
|
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::::'''Shewy Transformants'''
 +
:The image immediately to the right of this text shows the results from our transformation of S. oneidensis with plasmids pBAD18s, pBAD30, and pjQ200 via Zymo transformation and electroporation.
 +
 +
:The contents of the plates are as follows(column indicates transformation type):
 +
 +
{| align="center" border="1"
 +
|'''Zymo(streaked)'''||'''Zymo(poured)'''||'''Electroporation'''
 +
|-
 +
|pBAD18s|||pBAD18s|||pBAD18s
 +
|-
 +
|pBAD30|||pBAD30|||pBAD30
 +
|-
 +
|pjQ200|||pjQ200|||pjQ200
 +
|}
 +
 +
:The "poured" and "streaked" designations refer to how the transformed Shewy were put onto the plate. When transformed Shewy were streaked onto the plate, 5 microliters were used. When transformed Shewy were poured onto the plate, 100 microliters were used. All electroporated Shewy were poured onto plates.
 +
 +
|[[Image:BU_shewy_zymo_electro.jpg]]
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|}
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 +
[[Image:BU_electro_pour_ecoli_pbad18s.jpg]]
[[Image:BU_electro_pour_ecoli_pbad18s.jpg]]
[[Image:BU_electro_pour_ecoli_pbad18s_flash.jpg]]
[[Image:BU_electro_pour_ecoli_pbad18s_flash.jpg]]

Revision as of 15:19, 20 July 2007

Introduction to Electroporation and Testing

During the weekend of 7/14/07, the BU iGEM team ran a test of Electroporation of plasmids pBAD18s, pBAD30, and pjQ200 into wildtype S. oneidensis. Plasmids pBAD18s and pBAD30 were recommended to us by Professor Gardner as possible vectors for mutated Shewy genes. pjQ200 was the plasmid our team had previously selected as a prime candidate for serving as a vector. In order to check (and in the case of pjQ200, double-check) that these plasmids are capable of direct transformation into Shewy, tests of Zymo transformation and electroporation were conducted. For both types of transformation, E. coli was used as a control to verify that our transformation procedures were conducted properly. The results are posted below in a series of images and a short analysis. Lastly, gentamycin plates were used to select for pjq200 transformants, while ampicillin plates were used to select for pBAD18s and pBAD30 transformants.

Results of Electroporation

E. Coli Transformants
The image immediately to the right of this text shows the results from our transformation of E.coli with plasmids pBAD18s, pBAD30, and pjQ200 via Zymo transformation and electroporation.
The contents of the plates are as follows(column indicates transformation type):
Zymo(poured)Zymo(streaked)Electroporation
pBAD18spBAD18spBAD18s
pBAD30pBAD30pBAD30
pjQ200pjQ200pjQ200
The "poured" and "streaked" designations refer to how the transformed E. coli were put onto the plate. When transformed E. coli were streaked onto the plate, 5 microliters were used. When transformed E. coli were poured onto the plate, 100 microliters were used. Electroporated E. coli were all poured onto plates.

BU ecoli zymo electro.jpg

Shewy Transformants
The image immediately to the right of this text shows the results from our transformation of S. oneidensis with plasmids pBAD18s, pBAD30, and pjQ200 via Zymo transformation and electroporation.
The contents of the plates are as follows(column indicates transformation type):
Zymo(streaked)Zymo(poured)Electroporation
pBAD18spBAD18spBAD18s
pBAD30pBAD30pBAD30
pjQ200pjQ200pjQ200
The "poured" and "streaked" designations refer to how the transformed Shewy were put onto the plate. When transformed Shewy were streaked onto the plate, 5 microliters were used. When transformed Shewy were poured onto the plate, 100 microliters were used. All electroporated Shewy were poured onto plates.
BU shewy zymo electro.jpg


BU electro pour ecoli pbad18s.jpg BU electro pour ecoli pbad18s flash.jpg BU electro pour ecoli pbad30.jpg BU electro pour ecoli pjq200.jpg

BU electro pour shewy.jpg BU electro pour shewy 2.jpg BU electro pour shewy pbad18s.jpg BU electro pour shewy pbad18s bigger.jpg BU electro pour shewy pjq200.jpg

BU shewy zymo electro.jpg

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