User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/03

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June 3, 2013

Absorbance and Fluorescence Data

Finish preparing thiol DNA, AuNP and ThT sample<br.> 2[ThT]:1[DNA]→ 2.2245μM ThT<br.> 10% of 250μL=25μL (within 10% allows for accurate concentration comparison)<br.> Total solution volume: 275μL<br.> (2.2245×10-6M)(275×10-6L)=6.11738×10-10moles ThT<br.> (6.11738×10-10moles)/(25×10-6L)=24.5μM ThT<br.> <br.> To make 24.5μM ThT:<br.> (120μM)V=(24.5μM)(25μL)<br.> V=5.1μL in 19.9μL PBS buffer<br.> <br.> Add 25μL of 24.5μM ThT to 250μL DNA/AuNP solution<br.> New concentrations:

  • thiol DNA- 1.011μM
  • AuNP- 13.486μM
  • ThT- 2.2245μM

<br.> Abs data thiol DNA, AuNP, MB 06032013.png <br.> <br.> Abs data thiol DNA, ThT 06032013.png <br.> <br.> Fluor data thiol DNA, AuNP, MB 06032013.png <br.> <br.> Fluor data thiol DNA, AuNP, ThT 06032013.png

Notes

Samples containing Molecular Beacon excited at 532nm<br.> Samples containing ThT excited at 440nm<br.>

Sample Preparation for FCS and Calculations