User:Nelson Augusto Berrocal/Notebook/WiFi Coli 2010 Wet Lab/2010/06/01

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June 01st, 2010
I transformated competent cells with the biopart BBa_K098010 containing Ho1 y PcyA. I also transformated the OmpR inducible promoter (BBa_R0082).

BBa_K098010 is in the plasmid pSB3K3, in the 11N coordenate of Plate 3 form Kit iGem 2009; it has Kanamycin resistance.

BBa_R0082, in the plasmid pSB1A2 with ampicillin resistance; it was in the coordenate 16K from plate 1 Kit iGem 2009.

DNA Extraction Protocol from iGem Kit.

I must add 10μL in a pipet to the plate and resuspend; then I extract 10μL and I put 10μL in an eppendorf sterile.

Transformation Protocol

1)Defrost competent cells in ice

2)Add to each tube with 100μL with competent cells 4μL of DNA and let one with cells alone as a control

3)Let it rest in ice during 20min

4)Give a heat shock during 1min to 42°C

5)Let the tubes resting in ice during 5min

6)Add cells to assay tubes with 1ml of liquid LB

7)Keep the tubes shaking to 37°C during 1hr to 37°C

8)Plate in petri boxes with the apropiate selective medium, add 200μL per box


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