IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Gold Bioremediation/2010/10/21

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Minipreps of golS + RBS, golB + RBS, and golT + RBS was conducted and each was eluted into 40 uL of H2O.

50 uL digestions of the following:

  • golS + RBS
  • golB + RBS
  • golT + RBS
  • pSBIC3
  • B0015

A gel was extracted of golS and golB.

The cell digestions were then ran on a gel and ligated. The ligations included:

  • golB + RBS (from gel extraction), B0015, pSBIC3
  • golS + RBS (from gel extraction), B0015, pSBIC3

Heat shock transformation was used to incorporate the plasmids into the E.coli.

An earlier miniprep was verified using PCR, which included:

  • golS + RBS
  • golB + RBS
  • golT + RBS
  • pgolS + RBS
  • pgolS + E0420
  • pgols + E0430
  • pgolS + GFP

Inoculations were made from the earlier cell transformation to be incubated overnight.