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

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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


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