User:Allison K. Alix/Notebook/Thesis Research/2013/07/16
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UVVis Analysis/Silica Coating Pt. 2  Main project page Previous entry Next entry 
To Do
Silica Coating Procedure1) Prepare the following solutions:
3) Centrifuge solutions at 4000 rpm for one hour 4) Resuspend particles in PBS to a final concentration of 5nM? DNA/AuNP Data AnalysisSee Image:2013 0715 AuNPDNA.xlsx for AuNP/DNA spectra as well as absorbance values and molar extinction coefficients used to calculate concentrations of each in final solutions. Calculating the surface area of 1 AuNP Surface Area of a sphere = 4πr^{2}, r = 5nm 4π(5x10^{9}m)^{2} = 3.14 x 10^{16}m^{2} = 314nm^{2} Calculating the # of AuNP in 240.7μL 29.67nM solution 29.67 x 10^{9}mol/L x 240 x 10^{6}L x 6.02x10^{23} AuNP/mol = 4.299 x 10^{12} AuNP Calculating the Total Surface Area Available for DNA Attachment 4.299 x 10^{12} AuNP x 3.14x10^{16}m^{2}/1 AuNP = 0.001349886 m^{2} Calculating the Number of Moles of DNA Available to Attach 2.225x10^{6}moles/L x 240.7x10^{6}L = 5.3555575 x 10^{10} moles DNA Calculating oligonucleotide density on each particle (theoretical) 5.3555575x10^{10}moles DNA/0.001349886m^{2} AuNP = 3.967x10^{7} moles DNA/m^{2} AuNP (OR 39.67pmol DNA/cm^{2} AuNP) Calculating oligonucleotide density on each particle (actual) Sample A1 moles DNA = (1.827x10^{7}M x 350 x 10^{6}L) = 6.40x10^{11} moles 6.40x10^{11} moles/0.001349886m^{2} AuNP = 4.74x10^{8}moles DNA/m^{2}AuNP Sample B moles DNA = 4.44x10^{8}M x 500 x 10^{6}L = 2.22x10^{11} moles 2.22x10^{11} moles/0.001349886m^{2} AuNP = 1.64x10^{8}moles DNA/m^{2}AuNP Sample A2 moles DNA = 1.42x10^{10} 1.42x10^{10}/0.001349886m^{2} AuNP = 1.05x10^{7}moles DNA/m^{2}AuNP
Sample A1 moles DNA = (1.827x10^{7}M x 350 x 10^{6}L) = 6.40x10^{11} moles % hybridized = (6.40x10^{11}/5.3555575 x 10^{10} moles DNA)x100% = 11.95% Sample B moles DNA = 4.44x10^{8}M x 500 x 10^{6}L = 2.22x10^{11} moles % hybridized = (2.22x10^{11} moles/5.3555575 x 10^{10} moles DNA)x100% = 4.14% Sample A2 moles DNA = 1.42x10^{10} % hybridized = (1.42x10^{10}/5.3555575 x 10^{10}) = 26.5%
