User:Allison K. Alix/Notebook/Thesis Research/2013/05/15
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Objectives
Procedures
Calculations: 10x10-6mol/L x 500x10-6L = 5x10-9 moles ThT currently in solution 20x10-6mol/L x 550x10-6L = 1.1x10-8 moles ThT that should be in solution 1.1x10-8 moles - 5x10-9 moles = 6x10-9 moles ThT that need to be added 6x10-9moles/50x10-6L = 1.2x10-4M
Preparing 0.12mM ThT 5mM (x) = (0.12mM)(500μL) x = 12μL in 488μL PBS buffer NEW Concentrations: ThT: 20μM thiol-DNA: (8.9μM)(500μL) = x (550μL) x = 8.09μM Part 1: Attaching AuNP to thiol-DNA/ThT 1) Add 250μL thiol-DNA/ThT solution to 250μL 4% TEA/100mM DTT solution NEW Concentrations: 10μM ThT, 4.045μM thiol DNA 2) React 10 minutes 3) Extract DTT using 4 2mL aliquots of ethyl acetate 4) Mix 232.4μL thiol-DNA/ThT with 232.4μL 29.67nM AuNP and 35.2μL 284mM sodium citrate buffer 5) React 10 minutes NEW concentrations: 5μM ThT, 2.0225μM thiol-DNA, 14.835nM AuNP, 20mM sodium citrate 6) Centrifuge and redisperse in HEPES buffer Data
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