IGEM:UNAM-Genomics-Mexico/2009/Notebook/iGEM 2011 UNAM-Genomics Mexico/2011/05/31

{| width="800"
 * style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]] iGEM Project name 1
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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

PBBRMCS-5 DNA Plasmid extraction and Digestion
PBBRMCS-5 was extracted using alcaline lysis and 15μL Plasmid DNA digested with XBA I and Hind III

PBBRMCS-5 Plasmid DNA extracttion
We used Miguel Angel Ramirez's Method without the steps 11, and 13 through 20.

Gel Electrophoresis
The gel was made with 100 mL of TAE 1X and 1 gr. of Agarose, to have a gel with 1% agarose.


 * 1st Lane -> 5 μL of Ladder (Fermentas)
 * 2nd Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (1)
 * 3rd Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (2)
 * 4th Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (3)
 * 5th Lane -> 5 μL PBBRMCS-5 DNA Plasmid, 2 μL loading DNA dye. (4)
 * 6th Lane -> 2 μL of Loading DNA Dye.
 * 7th Lane -> 5 μL of Ladder (Fermentas)

The gel ran for 50 minutes at 110 volts

PBBRMCS-5 Plasmid Digestion

 * 2.5 μL of H20
 * 2.5 μL of NEBuffer 2 10X
 * 1.25 μL of XBA I
 * 1.25 μL of Hind III
 * 2.5 μL of BSA
 * 15 μL of PBBRMCS-5 Plasmid DNA

For a total reaction of 25 μL.

Leave all night at 37ºC in agitation.


 * }