User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/03
Measurements and Sample Prep for FCS | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
June 3, 2013Absorbance and Fluorescence DataFinish preparing thiol DNA, AuNP and ThT sample2[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:
<br.> Absorbance and Fluorescence Data<br.> <br.> <br.> <br.> <br.> <br.> NotesSamples containing Molecular Beacon excited at 532nm<br.> Samples containing ThT excited at 440nm<br.> Sample Preparation for FCS and CalculationsThiol DNA, MB, AuNP Solution Dilutions
1. 1nM AuNPs<br.> (9.03nM)V=(1nM)(30μL)<br.> V=3.32μL in 26.68μL PBS buffer
2. 500pM AuNPs<br.> 15μL 1nM solution in 15μL PBS buffer
3. 100pM AuNPs<br.> 6μL 500pM solution in 24μL PBS buffer
Thiol DNA, MB Solution Dilutions
4. 140.33nM MB<br.> 3.32μL in 26.68μL PBS buffer
5. 70.165nM MB<br.> 15μL 140.33nM MB in 15μL PBS buffer
6. 14.033nM MB<br.> 6μL 70.165nM MB in 24μL PBS buffer
MB Solution Dilutions
7. 140.33nM MB<br.> 3.32μL in 26.68μL PBS buffer<br.> <br.> 8. 70.165nM MB<br.> 15μL 140.33nM MB in 15μL PBS buffer<br.> <br.> 9. 14.033nM MB<br.> 6μL 70.165nM MB in 24μL PBS buffer<br.>
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