User:Tamra L. Fisher/Notebook/Experimental Biological Chem/2012/02/29
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To analyze the data from the UV-Vis and fluorescence readings for GFP from yesterday. To prepare for expression of MBP-intein.
The cultures from last night were spun down, resuspended, and transferred to the larger 1L LB cultures. Four hours later there was too much growth, and it was realized that the ampicillin was not added to the larger cultures, so they were now contaminated. The cultures were thrown out and a new expression will be completed next week.
Prep for MBP-intein expression
Data Analysis from Yesterday
Spectrum for GFP corrected with binding buffer spectrum:
A=εbc, where A485=0.165, b=1cm, and ε=61000cm-1M-1 The diluted concentration is calculated as 2.7μM. The concentration of the stock GFP is 27μM.
Fluorescence of 10,000 times dilute GFP was taken. The protein was excited at 485nm, and an emmision scan was taken from 490nm - 700.5nm: