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Team:Freiburg/Notebook/10 September
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| + | <a href="https://2011.igem.org/Team:Freiburg/Notebook/9_September">Previous entry</a> | ||
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| + | <a href="https://2011.igem.org/Team:Freiburg/Notebook"> 10 September </a> | ||
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==<span style="color:grey;">commons</span>== | ==<span style="color:grey;">commons</span>== | ||
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| + | 3. Transformation | ||
| style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:none;padding:0.0194in;"| insert 1 | | style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:none;padding:0.0194in;"| insert 1 | ||
| style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:0.0139in solid #808080;padding:0.0194in;"| 4 μl PcpcG from PCR | | style="border-top:none;border-bottom:0.0139in solid #808080;border-left:0.0139in solid #808080;border-right:0.0139in solid #808080;padding:0.0194in;"| 4 μl PcpcG from PCR | ||
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Transformation with the PR-♥-NOT-Ligation (6 different PR-vectors) | Transformation with the PR-♥-NOT-Ligation (6 different PR-vectors) | ||
| - | ==<span style="color: | + | ==<span style="color:orange;">Lysis cassette</span>== |
| - | === | + | ===M48 in C3 Vector=== |
| - | '''Investigators: | + | '''Investigators:Theo''' |
| + | Part M48 was Minipreped and cut with EcoRI and PstI. pSB1C3 was also PCR amplified on Friday (this was probably the problem for the failed Transformation of the other part-project -see Notebook Sept 9th-) and cut with EcoRI and PstI, ligated with M48 and transformed into competent cells. The colonies were to be picked on Sunday. | ||
| + | <br> | ||
| - | == | + | ===Troubleshooting of lysis cassette=== |
| - | + | ||
| - | = | + | |
| - | + | ||
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| + | Colonies of the transformation done on Friday were picked and inoculated on Amp LB medium | ||
==<span style="color:grey;">Precipitator</span>== | ==<span style="color:grey;">Precipitator</span>== | ||
Latest revision as of 00:23, 22 September 2011
Contact 
Contents |
commons
testdigest
Investigators:Julia
green light receptor
ligation for cloning PcpcG infront of CFP/YFP
Ligation
1. add H2O 11 μl
2. add 2 μl Ligase Buffer 10x
3. add Insert 1
4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
5. Add 1 μl T4-DNA Ligase
6. Incubate 30 min at room temperature
7. heat for 20 minutes at 80°C
| Name of part | |
| insert 1 | 4 μl PcpcG from PCR |
| vector | 2 μl [http://partsregistry.org/wiki/index.php?title=Part:BBa_I15016 BBa_I15016] /[http://partsregistry.org/wiki/index.php?title=Part:BBa_I15016 BBa_I1501]7 |
| H2O | 11 μl |
blue light receptor
Picking colonies
Investigators:Sophie
inoculating cm-LB with clones from the Gibson-♥-NOT-Trafo.
Labelled ♥-NOT-G a,b,c,d,e,f
Transformation
Investigators: Sophie
Transformation with the PR-♥-NOT-Ligation (6 different PR-vectors)
Lysis cassette
M48 in C3 Vector
Investigators:Theo
Part M48 was Minipreped and cut with EcoRI and PstI. pSB1C3 was also PCR amplified on Friday (this was probably the problem for the failed Transformation of the other part-project -see Notebook Sept 9th-) and cut with EcoRI and PstI, ligated with M48 and transformed into competent cells. The colonies were to be picked on Sunday.
Troubleshooting of lysis cassette
Colonies of the transformation done on Friday were picked and inoculated on Amp LB medium
Precipitator
Inoculation with pbd-clones
Investigators: Sophie
I inoculated 200mL Cm-LB with clones containing the plastic binding domain. I want to extract the GFP-pbd protein to be able to make experiments with this protein and get data about the plastic binding performance
