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Team:Freiburg/Notebook/3 September
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{{:Team:Freiburg/Templates/header}} | {{:Team:Freiburg/Templates/header}} | ||
| - | + | <html> | |
| + | <div id="notebook-page-header"> | ||
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| + | <a href="https://2011.igem.org/Team:Freiburg/Notebook/2_September">Previous entry</a> | ||
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| + | <div id="notebook-title"> | ||
| + | <a href="https://2011.igem.org/Team:Freiburg/Notebook"> 3 September </a> | ||
| + | </div> | ||
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| + | <a href="https://2011.igem.org/Team:Freiburg/Notebook/4_September">Next entry</a> | ||
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==<span style="color:green;">green light receptor</span>== | ==<span style="color:green;">green light receptor</span>== | ||
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'''Investigators: Julia''' | '''Investigators: Julia''' | ||
<br/> | <br/> | ||
| - | Continuing from | + | Continuing from 31 of Aug. [https://2011.igem.org/Team:Freiburg/Notebook/31_August]<br/> |
| - | Before Miniprep of 20 | + | Before Miniprep of 20 different colonies.<br/> |
Each minipreped DNA-sample (3μl)was digested with EcoRI and PstI.<br/> | Each minipreped DNA-sample (3μl)was digested with EcoRI and PstI.<br/> | ||
<br/> | <br/> | ||
| Line 14: | Line 26: | ||
<br/> | <br/> | ||
[[File:Freiburg11 09 03Testdigest 2A.Jpg|400px|400px]] | [[File:Freiburg11 09 03Testdigest 2A.Jpg|400px|400px]] | ||
| + | <br/> | ||
| + | |||
| + | |||
| + | |||
| + | the samples | ||
| + | |||
| + | 46b= ho1 with a RBS (BBa_B0034)<br/> | ||
| + | |||
| + | 50b= RBS(BBa_B0034)+pcyA+terminator<br/> | ||
| + | |||
| + | 64b= Promotor (BBa_J23110)+CcaS<br/> | ||
| + | |||
| + | 67b= Promotor (BBa_J23110)+CcaR<br/> | ||
| + | |||
| + | were send on [https://2011.igem.org/Team:Freiburg/Notebook/6_September 6.09] to GATC for sequencing | ||
===3A-assembly:Digestion=== | ===3A-assembly:Digestion=== | ||
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'''InsertI:''' | '''InsertI:''' | ||
| - | *64 | + | *64, CcaR |
| - | *46 | + | *46, ho1 |
with EcorI and SpeI | with EcorI and SpeI | ||
'''InsertII:''' | '''InsertII:''' | ||
| - | *67 | + | *67, CcaS |
| - | *50 | + | *50, pcyA |
with XbaI and PstI | with XbaI and PstI | ||
| - | Incubation for 6 hours at 37°C. Vector | + | For this assembly we used the constructs from the 2A-assembly made on [https://2011.igem.org/Team:Freiburg/Notebook/31_August 31.08] |
| + | |||
| + | Incubation for 6 hours at 37°C. Vector pSB1C3 was already digested,<br/> it was just treated with antarctic phosphatase for one hour. | ||
| + | |||
| + | ===3A-assembly:Ligation=== | ||
| + | |||
| + | '''Investigators: Julia''' | ||
==<span style="color:blue;">blue light receptor</span>== | ==<span style="color:blue;">blue light receptor</span>== | ||
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To do: Trafo | To do: Trafo | ||
| + | |||
| + | |||
| + | ===Transformation=== | ||
| + | with ♥C3 Lig. and Not C3 Lig | ||
| + | |||
| + | '''Investigator: Sophie''' | ||
==<span style="color:red;">red light receptor</span>== | ==<span style="color:red;">red light receptor</span>== | ||
Latest revision as of 00:27, 22 September 2011
Contact 
Contents |
green light receptor
Testdigestion of 2a-assembly
Investigators: Julia
Continuing from 31 of Aug. [1]
Before Miniprep of 20 different colonies.
Each minipreped DNA-sample (3μl)was digested with EcoRI and PstI.
Result:
the samples
46b= ho1 with a RBS (BBa_B0034)
50b= RBS(BBa_B0034)+pcyA+terminator
64b= Promotor (BBa_J23110)+CcaS
67b= Promotor (BBa_J23110)+CcaR
were send on 6.09 to GATC for sequencing
3A-assembly:Digestion
Investigators: Sandra
Digestion of:
InsertI:
- 64, CcaR
- 46, ho1
with EcorI and SpeI
InsertII:
- 67, CcaS
- 50, pcyA
with XbaI and PstI
For this assembly we used the constructs from the 2A-assembly made on 31.08
Incubation for 6 hours at 37°C. Vector pSB1C3 was already digested,
it was just treated with antarctic phosphatase for one hour.
3A-assembly:Ligation
Investigators: Julia
blue light receptor
Stop of ligation
Investigators: Sandra
Inactivation of ligase at 80°C for 20 minutes.
Stored in the blue light box, labelled: ♥C3 Lig., and Not C3 Lig.
To do: Trafo
Transformation
with ♥C3 Lig. and Not C3 Lig
Investigator: Sophie
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
Picking clones
Investigators: Sandra
Only few colonies grew on two plates. These clones were picked and incubated in LB with amp at 37°C.
