User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/04/02

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April 2, 2013

Hybridize thiol DNA and Molecular Beacon, then AuNP

DNA: 53.1μM → 727.5nM
V=13.7μL 53.1μM DNA in 486.3μL filtered H2O
V=100μL 1455nM DNA in 100μL H2O

MB: 7.31μM → 800nM

V=5.61μL 7.31μM MB in 44.53μL H2O

Combine 50μL 727.5nM thiol DNA with 50μL 800nM MB, heat at 75°C for 25 minutes, cool to room temperature.

Hybridize thiol DNA and ThT, then AuNP

ThT: from January 29, 2013- assumed 100%, Dr. Miller's 5mM ThT used in this procedure

V= 10μL of 5mM ThT in 9990μL PBS buffer
V= 160μL 5μM ThT in 840μL PBS buffer

Combine 50μL 727.5nM thiol DNA with 50μL 800nM ThT

AuNP with Ethanethiol

AuNP radius: (from Dr. Miller March 1, 2013) 14nm diameter=7nm radius
Area of AuNP: 4πr2=615.75 nm2
Sulfur atom radius: 200pm diameter=100pm radius
Area of sulfur atom: r2=40000pm2
sulfur atoms/AuNP=(6.1575×10-16m2)/(4×10-20m2)=15393

AuNP: 19.4nM
Ethanethiol: make 298μM, density = 0.839g/mL

2.5μL × (10-6L/1μL) × (1000mL/1L) × (0.839g/1mL) × (1 mol/ 62.134g) = 3.376×10-4mol = 0.34 M = 340 mM


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