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Team:Paris Saclay/Notebook/September/3
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[[File:Paris Saclay 3_september.jpg|250px|center]] | [[File:Paris Saclay 3_september.jpg|250px|center]] | ||
| - | [http://iramis.cea.fr/llb/Phocea/Pisp/visu.php?id=35&uid=%20alexei.grinbaum Alexei Grinbaum] , one of those few we have interviewed. | + | [http://iramis.cea.fr/llb/Phocea/Pisp/visu.php?id=35&uid=%20alexei.grinbaum Alexei Grinbaum] , one of those few we have interviewed during this month of september. |
| + | |||
| + | To learn more about the ethic aspect of our project, just click [https://2014.igem.org/Team:Paris_Saclay/Ethics here] | ||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Revision as of 00:28, 14 October 2014
Contents |
Wednesday 3rd September
LabWork
B- Construction of the fusion protein
We made a classic exttraction of plasmids from the liquid cultures.
Check by electrophoresis if it worked and send it for sequencing.
D- Lemon scent
by Mélanie
PCR LS
Electrophoresis of the PCR made Yesterday
well 2-3 = PCR with Dream taq or Vent taq
(well 4-5 = checking of the pps5 Extraction)
The PCR have success so I use the PCR clean up kit to purify it
Electrophoresis after purification
Digestion
In order to clone LS in pPSI, i Digest it by PacI
| component | volume |
|---|---|
| H2O | 3μl |
| buffer | 1μl |
| PacI | 1μl |
| PCR purify | 5μl |
Ligation
We already have some pPSI digested and dephosphorelated
So I do a ligation:
| component | volume |
|---|---|
| H2O | 5μl |
| buffer | 2μl |
| ligase | 1μl |
| pPSI | 2μl |
| LS PCR | 10μl |
2 hours at room temperature and over night at 4°
pPS5
Plasmid exctraction using the kit (picture is here)
digestion by SalI
| component | volume |
|---|---|
| H2O | 11μl |
| buffer | 5μl |
| SalI | 2μl |
| pPS5 | 30μl |
Electrophoresis
We saw that we have something strang with our plasmid but we wil check it tomorrow with a good ladder (the ladder here was normally on the well 1)
pPS3 and pPS4
We digest the plasmid by HindIII to direct our insert
| component | volume |
|---|---|
| H2O | 6μl |
| buffer | 1μl |
| HindIII | 1μl |
| pPS3/4 | 2μl |
well 1 = ladder well 2-3-4 = pPS3 Well 5-6-7 = pPS4
results are very strange
Photo of the Day
[http://iramis.cea.fr/llb/Phocea/Pisp/visu.php?id=35&uid=%20alexei.grinbaum Alexei Grinbaum] , one of those few we have interviewed during this month of september.
To learn more about the ethic aspect of our project, just click here
