User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/03/19

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March 19, 2013

Hybridization of thiol DNA, AuNP, and Molecular Beacon

DNA:AuNP ratio 75:1 AuNP concentration: 19.4nM thiol DNA concentration: 1455nM

  1. 100μL 5.5μM ThT, 100μL 727.5nM thiol DNA, and 9.7nM AuNP

heated for approximately 25 minutes at 70°C; cooled to room temperature

  1. Molecular Beacon concentration: 7.31μM = 7310nM

(7310)V=(800nM)(150μL) V= 5.5μL MB in 44.5μL filtered H2O Combine: 50μL 800nM MB, 50μL 727.5nM thiol DNA with 9.7nM AuNP heated for approximately 25 minutes at 70°C; cooled to room temperature

Procedure

thiol DNA and AuNP solution- 1. 500μL 2910nM DNA and 500μL 4% TEA with 100nM DTT; allowed to react for 10 minutes 2. 16mL (in 4mL aliquots) ethyl acetate used to extract (done under the fume hood) 3. 464.8μL 727.5nM DNA combined with 464.8μL 9.7nM AuNP and 70.4μL sodium citrate buffer; allowed to react for 10 minutes 4. centrifuge for 15-25 minutes



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