IGEM:Virginia United/2009/Notebook/2010 VT Lab/2010/07/07

From OpenWetWare
Jump to: navigation, search
Igem-logo-150px.png iGEM Project name 1 Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

July 7, 2010

  • Mini-prepped CFP (well 6A)
  • Quantified CFP plasmid DNA by using Nano Drop
    CFP – A	105.7 ng/ul
    CFP – B	56 ng/ul
  • Read the optical densities of the different solutions of copper with the TOP10 cells
    Incubated overnight with copper- 10¯³ 	1.4083 abs at 600 nm
    Incubated overnight with copper- 10¯³	1.4050
    Incubated overnight with copper- 10¯⁵	1.2855
    Incubated overnight with copper- 10¯⁵	1.1185
    Introduced with copper- 10¯³	        0.9670
    Introduced with copper- 10¯⁵   	        1.1208
    Introduced with copper- 10¯⁵   	        1.3408
    Top10 with no copper	                1.2049
  • Used Julie’s template and the CFP mini prepped in the morning and started the first round of PCR for constructing of the copper promoter
  • Analyzed fluorescence using plate reader of assorted cultures of CFP, YFP, RFP, and Top10 cells

Results: 10:45am – rfp flask fluorescing

12:50pm – Visual Fluorescence (strongest to weakest): rfp flask, rfp 1mL, rfp 5mL in 50mL

Plate Reader: rfp 1mL in 15mL and rfp5mL in 50mL. Both have the same OD But the 1mL has 2x the fluorescence

  • Figured out how to construct USER primers
  • Had a meeting with the Synthetic Biology team in the morning to discuss out current progress
  • Observed transformation results (Top10 cells with RFP biobricks with tetracycline resistance)
  • Emailed Wayne Outten and requested E.coli strains with knocked out genes for copper efflux
  • Consulted with Julie about PCR protocol
  • Discussed USER primer construction with Julie