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.

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 - P4 - T5 - o - P6

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

1. ladder 10µl
2. PCR result of CAD

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