User:Allison K. Alix/Notebook/Thesis Research/2013/06/05: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Silica Procedure Prep</span> | ||
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* 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 | '''Part 1: Preparing solutions for coating AuNP with Silica''' | ||
1) dissolve 0.716g PVP (10kg/mol) in 10mL water (0.00716M) | 1) dissolve 0.716g PVP (10kg/mol) in 10mL water (0.00716M) | ||
2) Dissolve 0.42mL NH<sub>4</sub>OH (29.3wt% NH<sub>3</sub> in H<sub>2</sub>O) in 9.58mL Ethanol (200 proof) (4.2vol%) | 2) Dissolve 0.42mL NH<sub>4</sub>OH (29.3wt% NH<sub>3</sub> in H<sub>2</sub>O) in 9.58mL Ethanol (200 proof) (4.2vol%) | ||
NOTE: TES solution will be prepared immediately prior to use to minimize exposure | NOTE: TES solution will be prepared immediately prior to use to minimize exposure | ||
Part 2: Preparing ZnPPIX solutions | |||
'''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 | |||
Revision as of 11:40, 12 July 2013
 Silica Procedure Prep
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Objectives
ProceduresPart 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
Zinc protoporphyrin (ZnPPIX) samples will be prepared at the same concentrations, following the same procedure as the ThT samples.
V=266.4μL 5μM ZnPPIX in 33.6μL (53.1μ)V=(2.2253μM)(300μL) V=12.57μL DNA in 287.43μL
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
(5000μM)V=(21.3μM)(25μL) V=0.107μL in 24.9μL
Concentrations: DNA- 0.94μM AuNP- 12.55nM ZnPPIX- 1.9364μM
V=133.2μL ZnPPIX in 166.8μL Data
NotesTo do:
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