User:Paulina Alatriste/Notebook/UNAM Genomics Mexico 2011/2011/06/24

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Plasmid isolation

ABSTRACT

  • Purification of pBBRMCS5 of BL21_DE3 cells.

  • Because of the plasmid that Pablo and I extracted yesterday was so dirty, today I isolate more. Pablo left cultures of BL21_DE3 cells to grow overnight. I follow the protocol of the following link: [[User:Paulina_Alatriste/Notebook/UNAM_Genomics_Mexico_2011/2011/06/14| Plasmid isolation].
  • Miguel told us that maybe the plasmid was dirty because the Solution III is not working properly so today I use another one, and also in step 5 after adding TE 10:1 I vortex the tubes to resuspend the pellet. I did an electrophoresis gel to see the quality of the plasmid:

Pato 24 jul 2011.png

  • Lanes 2-5 are the isolated plasmid. As you see is again dirty with chromosome residues so on Monday we will extract more using a Kit.