IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/07/15

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Continuing conjugation

I did the following steps with both of the stranis: DH5α and Bl21:

  • I took all the bacteria which grown overnigth and I put them into a test tube containing 1mL of MgSO4 and mixed with vortex.
  • I took 100 μL of the previous test and passed them to a test tube containing 900μL of MgSO4.(First dilution). I repeted this until reached third dilution.
  • I plaqued 100 μL of the second an thir dilution in a selective Py medium, contaning Nal an Gm because R. etli is natural resitant to Nal and if R. etli is correctly cojugated will aquire Gm resistance through pBBR1MCS-5.
  • The cultives will be left 2 days.

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