Team:Freiburg/Notebook/19 September
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+ | {{: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/18_September">Previous entry</a> | ||
+ | </div> | ||
+ | <div id="notebook-title"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook"> 19 September </a> | ||
+ | </div> | ||
+ | <div id="notebook-next"> | ||
+ | <a href="https://2011.igem.org/Team:Freiburg/Notebook/20_September">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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==<span style="color:orange;">Lysis cassette</span>== | ==<span style="color:orange;">Lysis cassette</span>== | ||
- | === | + | ===The_little_hoped_for_experiment_IV=== |
- | '''Investigators: | + | '''Investigators:Theo''' |
+ | I took the 3 cultures and prepared some tubes for them to test... I made measurements of the OD after about 18h of incubation at 30°C, put 1ml from every culture in an eppi (2 eppis for every culture) and incubated 3 of them at 30°C and 3 at 42°C. | ||
+ | The results can be seen on our Project Results page or the Registry (Part BBa_K608351 and BBa_K608352). | ||
+ | |||
+ | After that, I sent the samples for sequencing hoping I would get the results the next day, but typical GATC decided to delay them until the 21st of September!!! | ||
==<span style="color:grey;">Precipitator</span>== | ==<span style="color:grey;">Precipitator</span>== | ||
- | === | + | ===Ligation=== |
+ | |||
+ | |||
+ | {| 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:19.08. | ||
+ | |||
+ | |- | ||
+ | | 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 Name | ||
+ | |||
+ | Experiment Digestion19.08. Rüdiger /Gelextraction (Theo) | ||
+ | |||
+ | |- | ||
+ | | 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''' | ||
+ | |||
+ | |||
+ | PCR tube: | ||
+ | |||
+ | total volume 20 μl | ||
+ | |||
+ | |||
+ | # add H<sub>2</sub>O (17 μl -X-Y-Z) | ||
+ | # add 2 μl Ligase Buffer 10x | ||
+ | # add Insert 1, Insert 2(when proceeding from 3A digestion use 2 μl of each) | ||
+ | # add Vector (20ng needed. When proceeding from 3A digestion use 2 μl) | ||
+ | # Add 1 μl T4-DNA Ligase | ||
+ | # Incubate 10-30 min at room temperature | ||
+ | # heat for 20 minutes at 80°C | ||
+ | # store at -20°C or directly proceed to transformation | ||
+ | |||
+ | |||
+ | {| style="border-spacing:0;" | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name of part | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Ratio Insert:Vector | ||
+ | |||
+ | <nowiki>= 3:1 or 1:1</nowiki> | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Volume (μl) | ||
+ | |||
+ | |- | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| X insert 1 | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| <center>01:03</center> | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 10 | ||
+ | |||
+ | |- | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Y insert 2 | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | ||
+ | |||
+ | |- | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Z vector | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2 | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2 | ||
+ | |||
+ | |- | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| H<sub>2</sub>O | ||
+ | | style="background-color:#bfbfbf;border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | ||
+ | | style="background-color:#bfbfbf;border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| | ||
+ | | style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 5 | ||
+ | |||
+ | |} | ||
+ | '''Documentation:''' | ||
+ | |||
+ | Why are you doing this experiment? Where are your parts stored? Name the parts for ligation 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 ligate the 3 Precipitator motifs into iGEM Vector pSB1C3 | ||
+ | |||
+ | |} | ||
+ | |||
+ | |||
+ | |||
+ | ===NAME OF YOUR EXPERIMENT=== | ||
- | '''Investigators: | + | '''Investigators: NAME''' |
+ | {{:Team:Freiburg/Templates/footer}} |
Latest revision as of 00:21, 22 September 2011