User:Jorge Arturo Zepeda/Notebook/iGem LCG-UNAM team 2010/2010/06/30

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Wednesday 30th June 2010

I checked the disgestions on an agarose gel to confirm the existence of the ligation products expected, but the gel looks the same!! The ligations aren’t the ones we expected, so I’ll have to do it all over again.

IMG 0272.JPG

1º and 8º lanes are the DNA ladders
2º and 3º lanes are cI plasmid + GFP # 1
4º and 5º are cI plasmid + GFP # 2
6º and 7º are cI-PCR + GFP + Plasmid 18
Clearly we can see no bands standing for the ligation products expected.

I also extracted plasmids 18, 19 and 30, by growing them first in 4 ml of liquid LB with 4ul of the respective antibiotic, and the extract the plasmid with the High Pure Plasmid Isolation Kit from Roche. Plasmid 19 was also replated in a petri box of LB and ampicillin.

To be sure there was actually plasmid, I checked them on an agarose gel as follows:

IMG 0278.JPG

1º lane is the ladder
2º is plasmid 18
3º is plasmid 19
4º is plasmid 30