User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/04/15

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April 15th, 2010

1. Run gel to verify extracted plasmids and double restriction:

Ladder 1kb -> 1ul
DNA dye -> 1.5ul
DNA -> 5ul
Run gel for 40 min at 90V, then 10 min at 80V
Lanes: 1) Molecular Weight Marker; 2,3,5) Unknown; 4) Plasmid 18 (P-18); 6,7,9) Unknown; 8) P18 restriction; 10) P17 (Mariana); 11) P18 (M); 12) P19 (M); 13) P30 (M); 14) P17 rest. (M); 15) P18 rest. (M); 16) P19 rest. (M); 17) P30 rest. (M).


2. Plasmids 17, 19 and 30 discarded because I got confused about their identity, only plasmid 18 used to do more restriction (and Mariana’s plasmids 17, 18 & 30). Now I used:

DNA -> 25ul x 4.5
Buffer 2 -> 4ul
EcoRI -> 2ul
PstI -> 2ul
BSA -> 1ul
HPLC -> 6ul
Total volume: 40ul
Incubate at 37ºC overnight.



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