Monday 21st July

Lab Work

A - The chassis coli Odor free

Results: Transformation of CaCl₂ supercompetent cells

by Romain

Strains transformed: MG1655, MG1655Z1. from Bacterial Cultures transformed the 18th July

Results: Nothing has grown.

Results: Transformation of DY330 via pJBEI6409

by Sean

Transformation performed on the 18th July

Conclusion: The increase in colony number is proportional to the increase in volume used. The control yielded nothing. The results are coherent.

E - Banana scent

Oligonucleotide's design

by Romain

For the banana scent, design of iPS91 and iPS92, the ilvE sequence to transformed the Leucin into α-cetoisocaproate (The first step).

Liquid Bacterial Culture

by Marie, Romain & Sean

• DY330 pJBEI6409 with 10µl Cm in 10ml LB (x2)
• BT340 Cm and Amp
• The new strains received

Electrophoresis

by Fabio (process A) and Mathieu (process B and C)

We used 2µl of DNA of the following Biobricks in a 1% Agarose Gel. The PCRs came from Mathias' manipulation made the 18th July.

Process A

1. BBa_J23119 Cl1
2. BBa_J23119 Cl2
3. BBa_J23106 Cl1
4. BBa_J23100 Cl2
5. PCR 1
6. PCR 2
7. PCR 3
8. PCR 4
9. PCR 5
10. PCR 6
11. PCR 7
12. PCR 8
13. PCR 9
14. PCR 10

Process B

1. BBa_J23100 Cl1
2. BBa_J23100 Cl2
3. BBa_J23106 Cl1
4. BBa_J23106 Cl2
5. BBa_J23114 Cl1
6. BBa_J23114 Cl2

Process C

Pooling and purifying PCR 9 and 10 from process A.

1. PCR purified products. IPS70 / IPS71 of pOsV230 (Apramycin)
2. BT 340 (plasmid's flipase)

Results:

• A: From 1 to 4: Success, DNAs have the expected size.
• A: From 5 to 12: Failure, No PCR products.
• A: Numbers 13 and 14: Success, PCR products have the expected size.
• B: All 6 extractions were successful.
• C: Number 1: successful concentration of pOsV230's PCR product.
• C: Number 2 had no migration.

Protocol

People there:

• Instructors and advisors: Solenne and Sylvie.
• Students: Arnaud, Fabio, Juliette, Mathieu, Marie, Pierre, Romain, Sean and Terry.

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