IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/06/23

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Isolating Plasmid

  • Pato and I did the isolation of pBBRMCS-5 from E. coli BL21_DE3, accordanly to this protocol
  • The results are shown in the following electrophoresis gel(120V, 35 min):
 Lanes:
 1. Ladder 10 Kb.
 2. Isolation 1 by me.
 3. Isolation 2 by me.
 4. Isolation 1 by Pato.
 5. Isolation 2 by Pato.

Plasmid 23 jul 2011.JPG

Digestion

  • Miguel told us that the isolation was too dirty with chromosomal DNA, and that for a good digestion we need to perform the isolation again. Nevertheless we did a test digestion to see if Apa1 is working, if the restriction enzyme does not function properly, that means the strain we are using has the dcm methylase.

Isolation(again)

In order to obtain clean plasmid we performed the isolation exactly as the first time, except that we used a different solution 3, which according to Miguel could be the problem.

Paulina Alatriste(Pato) is gonna do it tomorrow.