IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Lock and Key Decoder/2010/08/10

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8/10

Plate

  • Spread Steve's 4 transformation recoveries onto plates.
  • Plate 1 - Plate 3 Tet? (Not sure, he got the plates out)
  • Plate 4 Amp? (Again, he got plates out)
  • Placed in 37°C room overnight.


Digestion
Reaction 1 Reaction 2 Reaction 3
RFP and Tet resistance backbone GFP (I13401) Downstream part Lock 1 (from strain 160)
500 ng DNA 500 ng DNA 500 ng DNA
5 μL NEB buffer 4 5 μL NEB buffer 4 5 μL NEB buffer 4
0.5 μL BSA 0.5 μL BSA 0.5 μL BSA
2 μL EcoR1 1 μL Xba1 1 μL EcoR1
2 μL Pst1 1 μL Pst1 1 μL Spe1
39.5 μL dH2O 41.5 μL dH2O 41.5 μL dH2O
  • 37°C bath for 1.25 hours
  • 80°C bath for 20 min.
  • Placed reaction tubes in -20°C freezer.
  • Matthew Entler 17:11, 10 August 2010 (EDT):



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