Friday 8th August

Lab work

C - Salicylate Inducible Suppressing System

Transformation of competent E.coli cells

by Fabio

We transformed in DH5α the Ligation's result made yesterday (BBa_K1372000 with BBa_B0015). The samples were put at 37°C for 10 hours, then we put them at room temperature until monday morning)

Samples:

1. Sample 2 μl
2. Sample 1 μl
3. DH5α Control in LB
4. DH5α Control in LB + Cm

Results on 11th August morning:

1. Success, grown very well (> 200 colonies)
2. Success, grown well (> 100 colonies)
3. Success, a film of bacteria has settled
4. Success, nothing has grown

Protocol

D - Lemon Scent

PCR

by Mélanie & Pierre We have done a PCR with pcola (geranyl synthase) and BBa_K517003:

• Green GotaqBuffer = 10µl
• MgCl2 = 4µl
• dNTP = 1µl
• Primer A = 1µM
• Primer B = 1µM
• DNA = 1µl
• Gotaq DNA polymerase = 0.25µl

Oligo used : iPS81bis AND iPS82 (pcola) iPS69 and iPS68bis (BBa_K517003)

PCR cycles:

• 95° 2min
• 95° 30sec
• 49-55° 30 sec
• 72° 2min
• 71° 5min

Electrophoresis

1. 10 µl of BBa_K517003
2. 10 µl of BBa_K517003
3. 10 µl of BBa_K517003
4. 10 µl of BBa_K517003
5. 10 µl of BBa_K517003
6. 10 µl of p. cola
7. 10 µl of p. cola
8. 10 µl of p. cola
9. 10 µl of p. cola
10. 10 µl of p. cola

Purification of the previous PCR

by Pierre

From the 5 tubes of p.Cola I cleaned up two tubes. From the 5 tubes of BBa_K517003 I cleaned up two tubes.

I did an electroforesis to check if the clening has worked. It did.

1. 1 µl of BBa_K517003
2. 1 µl of p. cola

Stock of pcola plasmides

using the DNA kit extraction to extract pcola ()

Human Practices

Art & Design

by Terry

Sequel of yesterday molding attempt.

The mold is cut delicately and the test piece removed.

Then, the mold is placed back in the becher and filled up with regular agar ( 20mg/ml )

Result : FAILURE, the agar test piece has been ripped when unmolding. It was stuck to the mold.

But the mold is still intact ! I cleaned it up and slightly covered the inward surface with liquid soap. The mold is filled up again and conserved in the refrigerator for 2 hours.

Result : SUCCESS, I slightly remoistened the gel with water for an easier unmolding. The agar test piece is perfectly removed.

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