IGEM:UNAM-Genomics Mexico/2009/Notebook/H2/2011/07/28

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Green Light Receptor

Goal: prepare two constructions for ligation. Results: in preparation

Construction 1

Contents: BBa_J23117(promoter) + RBS + CcaR + CcaS; pSB1A3 (ampicilin)

Work done: duplicate plasmid extraction with Kit; duplicate plasmid restriction with XbaI & PstI.

Restriction mixture (40μL total):

  1. 15μL DNA
  2. 1μL BSA
  3. 4μL Buffer #2
  4. 2μL Enzyme PstI
  5. 2μL Enzyme XbaI
  6. 16μL H2O

Construction 2

Contents: PcyA + Ho1; pSB?K? (kanamycin)

Work done: culture undergoing re-growth as colonies were too small.

Further work

Digest construction 2 and prepare for insertion.

Determine best outcome, insert C2 into C1's plamid or vice versa.

Ligate construction 1 + construction 2 + BBa_b0015(dual terminator)


Modeling

Model plasmid stability as a continuous Markov model. I got a good reference from a teacher on how to go from the transition matrix to the expected value (Martínez et al. Paper on the Believable Ending of the Twilight Saga. Really, not kidding).


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