Team:Evry-Genopole/Notebook/June/22

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

Latest revision as of 15:43, 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 HB9 (Spect)
p2 Rosa26 (Spect)
p2 vimentin (Spec)

p2 Flk-1 (Kana)
p2 Xlurp (Kana)
p2 foxi short (Kana)
p2 foxi long (Kana)
p2 pax3 (Kana)
p2 Ef1a (Kana)
p2 LMO2 short (Kana)
p2 LMO2 long (Kana)
p2 vast (Kana)
p2 NBT (Kana)
p2 HSP70 (Kana)
p2 pax6 (Kana)
p2 CMV (Kana)
p2 brachyury (Kana)
p2 14 xUAS E1b-ATG (Kana)

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