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From 2014.igem.org

Contents 1 Monday 21st July 1.1 Lab Work 1.1.1 1 - Results: Transformation of supercompetent cells with CaCl2 1.1.2 2 - Results: Transformation of DY330 via pJBEI6409 1.1.3 3 - Oligo's design 1.1.4 4 - Liquid Bacterial Culture 1.1.5 5 - Electrophoresis

Monday 21st July

Lab Work

1 - Results: Transformation of supercompetent cells with CaCl₂

by Romain

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

Results: Nothing has grown.

2 - Results: Transformation of DY330 via pJBEI6409

by Sean

Transformation performed on the 18th July

Number of colonies per dish

50μL from cuvette	100μL	remainder
0	4	30	2μL of plasmid
3	6	45	4μL
0	0	0	Control

3 - Oligo's design

by Romain

4 - Liquid Bacterial Culture

by Marie, Romain & Sean

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

5 - 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. J23119 Cl1
2. J23119 Cl2
3. J23106 Cl1
4. 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. J23100 Cl1
2. J23100 Cl2
3. J23106 Cl1
4. J23106 Cl2
5. J23114 Cl1
6. 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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