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

{| width="800"
 * style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]] iGEM UNAM-Genomics-Mexico
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

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.


 * }