User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/05/31

Measurements and Sample Prep

Main project page Previous entry      Next entry

May 31, 2013 Absorbance and Florescence Measurements <br.> <br.> <br.> Concentrations thiol DNA- 615.59nM<br.> AuNP- 9.03nM<br.> Molecular Beacon- 1268nM<br.> Notes Samples excited at 532nm, fluorescence peak observed at approximately 565nm Sample Preparation Molecular Beacon, final concentration: 1268nM (7.61μM)V=(1.268μM)(100μL) V=16.7μL 7.61μM MB in 83.3μL PBS buffer AuNP and thiol DNA, final concentrations: 14.835μM and 1.11265μM (53.1μM)V=(4.45μM)(250μL)<br.> V=20.95μL 53.1μM thiol DNA in 229.05μL PBS buffer<br.> thiol DNA extraction following the same method from Allison Alix on 05/15/2013 without ThT Combine 232μL 29.67nM, 10nm AuNP, 232μL extracted thiol DNA, and 36μL sodium citrate buffer Always add sodium citrate buffer last, to avoid disruption of DNA by the change in pH To be continued on Monday, June 3, 2013 Try to sonicate AuNPs ThT and thiol DNA, final concentrations: 2.2244μM and 1.011μM (53.1μM)V=(2.022μM)(250μL)<br.> V=55.6μL 53.1μM thiol DNA in 194.38μL PBS buffer<br.> (20μM)V=(4.45μM)(250μL)<br.> V=55.6μL 20μM ThT in 194.38μL PBS buffer<br.> ThT, final concentration: 2.2245μM (20μM)V=(2.2245μM)(250μL)<br.> V=27.8μL 20μM ThT in 222.2μL PBS buffer<br.> Notes Finish preparing AuNP and thiol DNA sample Run absorbance and fluorescence for all four samples Compile data into one absorbance graph and one fluorescence graph Eventually run samples on FCS Next week: FCS, silica-coating on nanoparticles