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Team:Paris Saclay/Notebook/August/25
From 2014.igem.org
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D : sample digested by XbaI and PstI | D : sample digested by XbaI and PstI | ||
| - | T1 - D1 - T2 - D2 - T3 - D3 - E - T4 - | + | T1 - D1 - T2 - D2 - T3 - D3 - E - T4 - D4 - T5 - o - D6 |
[[File:250814 laetitia digestion verif chromoP pGEMTeasy.jpg|500px|]] | [[File:250814 laetitia digestion verif chromoP pGEMTeasy.jpg|500px|]] | ||
Revision as of 14:52, 25 August 2014
Contents |
Monday 25th August
Lab work
B - Construction of the fusion protein
plasmid extraction : pGEMTeasy + chromoprotein
by Laetitia
We used the bacteria containing pGEMTeasy + chromoprotein launched by Melanie sunday the 24th for the plasmid extraction, 6 independent cultures which come from 6 independent colonies.
We made a stock of bacteria for each sample (x6).
To extract the plasmid, we used the plasmid DNA purification kit (Macherey-Nagel).
Digestion of pGEMTeasy+chromoproteinby XbaI and PstI
by Laetitia
The goal is to check the presence of the insert inside the plasmid. We digest the 6 samples of purified plasmid.
| component | volume |
|---|---|
| Plasmid | 5 μl |
| Fast Digest buffer 10X | 1μl |
| XbaI | 0.5μl |
| PstI | 0.5μl |
| H20 | 3μl |
1h at 37°C
Electrophoresis of the digestion product of pGEMTeasy+chromoprotein
by Laetitia
T : control negatif without enzyme
D : sample digested by XbaI and PstI
T1 - D1 - T2 - D2 - T3 - D3 - E - T4 - D4 - T5 - o - D6
Results : the samples 3, 4 and 6 seems to contain the chromoprotein gene (two bands)
Bacterial culture of DH5a containing pGEMTeasy + chromprotein
by Laetitia
We launched 6 cultures in 5mL LB + Ampi (1/1000) - 37°C at night . The bacteria come from the stock made before the plasmid extraction
D - Lemon Scent
Gel electrophoresis of CAD
by Sean
Legend
- ladder 10µl
- PCR result of CAD
