Team:Evry-Genopole/Notebook/June/22

From 2012.igem.org

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Bacterial Transformation
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{{:Team:Evry-Genopole/template_v1}}
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Bacteria: T10
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<html>
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DNA: pCS2 (+) at 640 ng.uL-1
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<center><h1>Promoters & Reporters workgroup</h1></center>
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</html>
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1) Keep constantly the cells on ice
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<FONT SIZE=3>'''Bacterial Transformation'''</FONT> <br/>
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2) Add 1 uL of pCS2 (+) in 100 uL of T10
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3) Incubate 30 min on ice
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Bacteria: T10<br/>
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4) Heat shock the cells during 30 sec at 42 degree Celsius in a water-bath without shaking
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DNA: pCS2 (+) at 640 ng.uL-1<br/>
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5) Put 2 min on ice
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6) Add 500 uL of pre warmed SOC-medium
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1) Keep constantly the cells on ice <br/>
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7) Incubate 1h at 37 degree Celsius at 225 rpm
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2) Add 1 uL of pCS2 (+) in 100 uL of T10<br/>
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8) Spin at 5000 rpm during 30 sec
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3) Incubate 30 min on ice<br/>
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9) Remove 150 uL - 400 uL of supernatant
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4) Heat shock the cells during 30 sec at 42 degree Celsius in a water-bath without shaking<br/>
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10) Resuspend the pellet in the 150 uL left
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5) Put 2 min on ice<br/>
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11) Spread on appropriate plates
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6) Add 500 uL of pre warmed SOC-medium<br/>
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12) Incubate overnight at 37 degree Celsius
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7) Incubate 1h at 37 degree Celsius at 225 rpm<br/>
 +
8) Spin at 5000 rpm during 30 sec<br/>
 +
9) Remove 150 uL - 400 uL of supernatant<br/>
 +
10) Resuspend the pellet in the 150 uL left<br/>
 +
11) Spread on appropriate plates<br/>
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12) Incubate overnight at 37 degree Celsius<br/>
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List of promoters received:
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List of promoters received:<br/>
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p2 Flk-1
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p2 Flk-1<br/>
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p2 Xlurp
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p2 Xlurp<br/>
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p2 foxi short
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p2 foxi short<br/>
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p2 foxi long
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p2 foxi long<br/>
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p2 pax3
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p2 pax3<br/>
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p2 Ef1a
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p2 Ef1a<br/>
-
p2 Rosa26
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p2 Rosa26<br/>
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p2 LMO2 short
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p2 LMO2 short<br/>
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p2 LMO2 long
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p2 LMO2 long<br/>
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p2 vast
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p2 vast<br/>
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p2 HB9
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p2 HB9<br/>
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p2 NBT
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p2 NBT<br/>
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p2 HSP70
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p2 HSP70<br/>
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p2 pax6
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p2 pax6<br/>
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p2 CMV
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p2 CMV<br/>
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p2 brachyury
+
p2 brachyury<br/>
-
p2 vimentin
+
p2 vimentin<br/>
-
p2 14 xUAS E1b-ATG
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p2 14 xUAS E1b-ATG<br/>

Revision as of 15:35, 29 June 2012

Team IGEM Evry 2012

Promoters & Reporters workgroup

Bacterial Transformation

Bacteria: T10
DNA: pCS2 (+) at 640 ng.uL-1

1) Keep constantly the cells on ice
2) Add 1 uL of pCS2 (+) in 100 uL of T10
3) Incubate 30 min on ice
4) Heat shock the cells during 30 sec at 42 degree Celsius in a water-bath without shaking
5) Put 2 min on ice
6) Add 500 uL of pre warmed SOC-medium
7) Incubate 1h at 37 degree Celsius at 225 rpm
8) Spin at 5000 rpm during 30 sec
9) Remove 150 uL - 400 uL of supernatant
10) Resuspend the pellet in the 150 uL left
11) Spread on appropriate plates
12) Incubate overnight at 37 degree Celsius

List of promoters received:

p2 Flk-1
p2 Xlurp
p2 foxi short
p2 foxi long
p2 pax3
p2 Ef1a
p2 Rosa26
p2 LMO2 short
p2 LMO2 long
p2 vast
p2 HB9
p2 NBT
p2 HSP70
p2 pax6
p2 CMV
p2 brachyury
p2 vimentin
p2 14 xUAS E1b-ATG

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