User:Nelson Augusto Berrocal/Notebook/WiFi Coli 2010 Wet Lab/2010/04/26

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April 26th, 2010

I made a restriction with EcoRI and XbaI to the last restriction of plasmid with the biobrick BBa_B0015. The restriction was made with 40μL, which were composed of:
20μL DNA
4μL Buffer 2
1μL each enzyme
1μL BSA
14μL H2O
To observe if the restriction and the plasmid were OK I made an agarose Gel to .08.
Ladder | DNA Restricted 4μL | Plasmid 5μL | Plasmid Paz(control) 5μL | Ladder
Gel 90V 40min + 80V 10min
The plasmid we were looking for is pSB1AK3 with 100-300 copy number per cell. 3189bp
We got as a result two fragments of 1kb(???) and 3kb instead the 3kb and 100bp fragments that we expected.