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

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{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]      }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]}} |style="background-color: #F2F2F2" align="center"| [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]
{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]      }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]}} |- |- Line 9: Line 9: * Prepare solutions for silica procedure (See: [[User:Allison K. Alix/Notebook/Thesis Research/2013/05/24]]) * 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 * Prepare ZnPPIX, ZnPPIX/DNA, and ZnPPIX/DNA/AuNP solutions + * Take UV/Vis and Fluorescence of ZnPPIX Samples ==Procedures== ==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== + + [[Image:Pic_of_ZnPPIX_abs.png]] + + [[Image:Pic_of_ZnPPIX_fluor.png]] + + ==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 + +

## Current revision

Silica Procedure Prep Main project page
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## 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

## 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