User:University of Washington/29 July 2008

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luxR from pLac

-Bacterial stab of part I0462 received from iGEM HQ.

-Sequence of R0010+E0240 ligation received back and found to be incorrect. Assembly of these parts must be retried.

-Restriction digests of R0010 and E0240 started; will incubate at 37 overnight.

LuxR from AraC and TetR

-PCR purified the AraC and TetR promoter.

-Digest the Elowitz's plasmid with enzyme HindIII to find out what plasmid exactly the MG1655Z1 strain contain. Two potential >> pCS26 has 4 sites: expected 4716, 1990, 1982, 785 bp and pACYC184 has 1 site: 4245 bp

  • 34.5ul ddH2O + 5ul NEB2 + 10ul DNA(assume 100 ng/ul) + ...vortex... + 0.5 ul HindIII +....centrifuge ...
  • incubated 37 degree Celsius for 1.5 hours

-Ran gel. Saw two bands, one between 4 and 5 kb, the other between 500 - 1000 bp. The control(no enzyme) didn't have small fractions.


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