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From 2012.igem.org

Weeks

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2

3

4

5

6

7

8

9

10

11

12

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16

Week 3: 25th June - 1st July

Monday, 25th June

Bacterial Transformation

Bacteria:T10
Reporters:

• RFP: P1-18F K
• GFP: P1-14K A
• CFP: P1-6A A
• YFP: P2/24E AK
• OFP: P2/13N K

• Violecein P3/12B T
• Vio operon ABDE P3/20H K
• Vio operon ABCE P3/20J K
• CelEBI w/RRS P3/6H A
• CelEBY w/RRS P3/6L H

Protocol:

1. Keep constantly the cells on ice
2. Rehydratation of DNA in 10uL H2O 2) Add 1 uL of DNA in 50 uL of T10
3. Incubate 30 min on ice
4. Heat shock the cells during 30 sec at 42 degree Celsius in a water-bath without shaking
5. Put 2 min on ice
6. Add 500 uL of pre warmed SOC-medium
7. Incubate 1h at 37 degree Celsius at 225 rpm
8. Spin at 5000 rpm during 30 sec
9. Remove 150 uL - 400 uL of supernatant
10. Resuspend the pellet in the 150 uL left
11. Spread on appropriate plates
12. Incubate overnight at 37 degree Celsius

Tuesday, 26th June

Wednesday, 27th June

Thursday, 28th June

Friday, 29th June