Team:Freiburg/Notebook/20 August
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{{:Team:Freiburg/Templates/header}} | {{: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/19_August">Previous entry</a> | ||
+ | </div> | ||
+ | <div id="notebook-title"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook"> 20 August </a> | ||
+ | </div> | ||
+ | <div id="notebook-next"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/21_August">Next entry</a> | ||
+ | </div> | ||
+ | </div> | ||
+ | </html> | ||
==<span style="color:green;">green light receptor</span>== | ==<span style="color:green;">green light receptor</span>== | ||
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continue from: Transformation, 19.08.11<br/> | continue from: Transformation, 19.08.11<br/> | ||
Testdigest showed, that cloning was not succesful. Seems that the vector was only partly digested, because only vector-bands were visible on gel. I will try it again with longer digestion times. | Testdigest showed, that cloning was not succesful. Seems that the vector was only partly digested, because only vector-bands were visible on gel. I will try it again with longer digestion times. | ||
+ | |||
+ | ===Miniprep=== | ||
+ | |||
+ | '''Investigators: Rüdiger''' | ||
+ | |||
+ | Miniprep of PET.DUET_1 vector. | ||
+ | |||
+ | ===Trafo=== | ||
+ | |||
+ | |||
+ | {| style="border-spacing:0;" | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name:Rüdiger | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date:20.08 | ||
+ | |||
+ | |- | ||
+ | | colspan="2" style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Date 19.08. Name Rüdiger | ||
+ | |||
+ | Experiment Ligation | ||
+ | |||
+ | |- | ||
+ | | colspan="2" style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name: Precipitator | ||
+ | |||
+ | |} | ||
+ | Procedure | ||
+ | |||
+ | |||
+ | # take cells from -80°C freezer and put them on ice! (every eppi contains about 400 μl cells) | ||
+ | # thaw cells on ice 20 minutes | ||
+ | # pipette 50 μl cells and 2 μl DNA into eppi still on ice! | ||
+ | # Incubate for 30 minutes on ice | ||
+ | # Heat at 42°C for 60 sec | ||
+ | # Incubate on ice for 5 minutes | ||
+ | # Add 200 μl LB Broth | ||
+ | # Incubate for 2 hours at 37°C (cells with lysis cassette at 30°C!!) | ||
+ | # Plate 50 μl and 200μl on two different LB/Agar plates with appropriate antibiotic resistance | ||
+ | |||
+ | '''Documentation:''' | ||
+ | |||
+ | Why are you doing this experiment? Name of the sample? Where are they stored? Name the vector with inserts, antibiotika resistance etc. | ||
+ | |||
+ | |||
+ | {| style="border-spacing:0;" | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Trying to get Precipitator into iGEM vector. | ||
+ | |||
+ | Did not work. | ||
+ | |||
+ | |} | ||
+ | |||
+ | |||
+ | |||
+ | '''Investigators: Rüdiger''' | ||
+ | |||
+ | Transformation of precipitator(pSB1C3). | ||
+ | |||
+ | Trafo did not work. | ||
+ | |||
+ | 10.09. | ||
+ | |||
+ | Rüdiger | ||
+ | |||
+ | |||
+ | Picked cells from yesterday's transformation | ||
+ | |||
+ | |||
+ | 1 a/b A-C | ||
+ | |||
+ | 2 a/b A-C | ||
+ | |||
+ | 3 a/b A-C | ||
+ | |||
+ | 5 a/b A-C | ||
+ | |||
+ | 6 a/b A-C | ||
+ | |||
+ | 7 a/b A-C | ||
+ | |||
+ | |||
+ | -miniprep of yesterday's inoculation. | ||
+ | |||
+ | |||
+ | 11.09. | ||
+ | |||
+ | -miniprep of yesterday's inoculation. | ||
+ | |||
+ | |||
{{:Team:Freiburg/Templates/footer}} | {{:Team:Freiburg/Templates/footer}} |
Latest revision as of 01:10, 22 September 2011