Team:Freiburg/Notebook/3 September
From 2011.igem.org
(Difference between revisions)
(Created page with "==<span style="color:green;">green light receptor</span>== ===NAME OF YOUR EXPERIMENT=== '''Investigators:NAME''' ==<span style="color:blue;">blue light receptor</span>== =...") |
|||
(19 intermediate revisions not shown) | |||
Line 1: | Line 1: | ||
+ | {{:Team:Freiburg/Templates/header}} | ||
+ | <html> | ||
+ | <div id="notebook-page-header"> | ||
+ | <div id="notebook-back" width="100px" > | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/2_September">Previous entry</a> | ||
+ | </div> | ||
+ | <div id="notebook-title"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook"> 3 September </a> | ||
+ | </div> | ||
+ | <div id="notebook-next"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/4_September">Next entry</a> | ||
+ | </div> | ||
+ | </div> | ||
+ | </html> | ||
==<span style="color:green;">green light receptor</span>== | ==<span style="color:green;">green light receptor</span>== | ||
- | === | + | ===Testdigestion of 2a-assembly=== |
- | '''Investigators: | + | '''Investigators: Julia''' |
+ | <br/> | ||
+ | Continuing from 31 of Aug. [https://2011.igem.org/Team:Freiburg/Notebook/31_August]<br/> | ||
+ | Before Miniprep of 20 different colonies.<br/> | ||
+ | Each minipreped DNA-sample (3μl)was digested with EcoRI and PstI.<br/> | ||
+ | <br/> | ||
+ | Result:<br/> | ||
+ | <br/> | ||
+ | [[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=== | ||
+ | |||
+ | '''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 [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>== | ||
- | === | + | ===Stop of ligation=== |
- | '''Investigators: | + | '''Investigators: Sandra''' |
+ | Inactivation of ligase at 80°C for 20 minutes.<br/> | ||
+ | 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''' | ||
==<span style="color:red;">red light receptor</span>== | ==<span style="color:red;">red light receptor</span>== | ||
Line 35: | Line 104: | ||
==<span style="color:grey;">Precipitator</span>== | ==<span style="color:grey;">Precipitator</span>== | ||
- | === | + | ===Picking clones=== |
+ | |||
+ | '''Investigators: Sandra''' | ||
+ | |||
+ | Only few colonies grew on two plates. These clones were picked and incubated in LB with amp at 37°C. | ||
- | + | {{:Team:Freiburg/Templates/footer}} |
Latest revision as of 00:27, 22 September 2011