User:Allison K. Alix/Notebook/Thesis Research/2013/06/05

Silica Procedure Prep

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Objectives Prepare solutions for silica procedure (See: User:Allison K. Alix/Notebook/Thesis Research/2013/05/24) Prepare ZnPPIX, ZnPPIX/DNA, and ZnPPIX/DNA/AuNP solutions Take UV/Vis and Fluorescence of ZnPPIX Samples Procedures Part 1: Preparing solutions for coating AuNP with Silica 1) dissolve 0.716g PVP (10kg/mol) in 10mL water (0.00716M) 2) Dissolve 0.42mL NH4OH (29.3wt% NH3 in H2O) in 9.58mL Ethanol (200 proof) (4.2vol%) NOTE: TES solution will be prepared immediately prior to use to minimize exposure Part 2: Preparing ZnPPIX solutions Zinc protoporphyrin (ZnPPIX) samples will be prepared at the same concentrations, following the same procedure as the ThT samples. 2.22μM ZnPPIX and 1.11265μM DNA (5μM)V=(4.44μM)(300μL) V=266.4μL 5μM ZnPPIX in 33.6μL (53.1μ)V=(2.2253μM)(300μL) V=12.57μL DNA in 287.43μL ZnPPIX, DNA and AuNP 2[ThT]:1[DNA]→ 2.068μM ZnPPIX 10% of 250μL=25μL (within 10% allows for accurate concentration comparison) Total solution volume: 275μL (2.068×10-6M)(275×10-6L)=5.31476×10-10moles ZnPPIX (5.31476×10-10moles)/(25×10-6L)=21.3μM ZnPPIX Make 21.3μM ZnPPIX: (5000μM)V=(21.3μM)(25μL) V=0.107μL in 24.9μL Add 25μL of 21.3μM ZnPPIX to 250μL DNA/AuNP solution Concentrations: DNA- 0.94μM AuNP- 12.55nM ZnPPIX- 1.9364μM 2.22μM ZnPPIX (5μM)V=(2.22μM)(300μL) V=133.2μL ZnPPIX in 166.8μL Data Notes To do: Prepare new ZnPPIX/DNA sample and take fluorescence (it is curious that there was no fluorescence seen in this sample) Prepare AuNP/ZnPPIX sample and take fluorescence (be sure that the fluorescence seen in the AuNP/DNA/ZnPPIX sample is not due to an AuNP/ZnPPIX interaction) Dilute beads 10X and 100X more, there was WAY too much signal and no curve seen in FCS measurements