User:Allison K. Alix/Notebook/Thesis Research/2013/08/13
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Scaled up Silica-Coating Procedure
Original: 1mL AuNP & MHA solution/2μL TES/0.125mL NH4OH/5mL 2-propanol
1) Re-scale to use 9mL AuNP/MHA solution
New: 9mL AuNP&MHA solution/18uL TES/1.125mL NH4OH/45mL 2-propanol
2) Stir solutions for one hour to allow reaction to proceed
3) Centrifuge solutions at 4000 rpm for one hour
4) Re-suspend particles in water
Original concentration of AuNP/MHA solution is 4.7nM
0.767x10^-9moles/L x 1L/1000mL x 36mL = 2.7612 x 10^-11 moles AuNP total
2.7612 x 10^-11 moles AuNP x 6.02 x 10^23 particles/mole = 1.66 x 10^13 total Au nanoparticles
5x10^-9 moles/L x 1L/1000mL x 6.02x10^23 particles/mole = 3.01 x 10^12 particles/mL
1.66 x 10^13 particles / x = 3.01 x10^12 particles/1 mL
x = 5.52mL
Dilute AuNP in 5.52mL water after removing supernantant
5.52mL/24 vials = 230uL per vial.
DNA Attachment Procedure 1
1) Mix 1mL 5nM AuNP@SiO2 with 5uL 1mM APS. Stir for 1hr.
2) Reduce thiol DNA to HS-DNA using DTT. Dissolve 2mg Sulfo-SMCC in 2mL 0.05M KP buffer. Mix 250uL solution with 250uL 3uM DNA. Mix for 1 hour
3) Centrifuge DNA/Sulfo-SMCC solution for 20 minutes at 13,200rpm. Remove excess Sulfo-SMCC and redisperse DNA in 250uL PBS.
4) Mix 250uL Sulfo-SMCC activated DNA with
DNA Attachment Procedure 2