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
2[ThT]:1[DNA]→ 2.2245μM ThT
10% of 250μL=25μL (within 10% allows for accurate concentration comparison)
Total solution volume: 275μL
(2.2245×10-6M)(275×10-6L)=6.11738×10-10moles ThT
(6.11738×10-10moles)/(25×10-6L)=24.5μM ThT

To make 24.5μM ThT:
(120μM)V=(24.5μM)(25μL)
V=5.1μL in 19.9μL PBS buffer

Add 25μL of 24.5μM ThT to 250μL DNA/AuNP solution
New concentrations:

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


Image:Abs_data_thiol_DNA,_AuNP,_MB_06032013.png

Image:Abs_data_thiol_DNA,_ThT_06032013.png

Image:Fluor_data_thiol_DNA,_AuNP,_MB_06032013.png

Image:Fluor_data_thiol_DNA,_AuNP,_ThT_06032013.png

Notes

Samples containing Molecular Beacon excited at 532nm
Samples containing ThT excited at 440nm

Sample Preparation for FCS and Calculations


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