User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/07/25: Difference between revisions
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*Day Two: | *Day Two: | ||
**Prepare the following solutions: | **Prepare the following solutions: | ||
#1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/5μL TES/0.125mL | #1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/5μL TES/0.125mL NH<sub>4</sub>OH/5mL isopropanol | ||
#1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/4μL TES/0.125mL | #1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/4μL TES/0.125mL NH<sub>4</sub>OH/5mL isopropanol | ||
#1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/3μL TES/0.125mL | #1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/3μL TES/0.125mL NH<sub>4</sub>OH/5mL isopropanol | ||
#1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/2μL TES/0.125mL | #1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/2μL TES/0.125mL NH<sub>4</sub>OH/5mL isopropanol | ||
#1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/1μL TES/0.125mL | #1mL 9.4nM, 9.65nm AuNP&MHA, MQ H<sub>2</sub>O/1μL TES/0.125mL NH<sub>4</sub>OH/5mL isopropanol | ||
#*Stir solutions for one hour to allow reaction to proceed | #*Stir solutions for one hour to allow reaction to proceed | ||
#*Centrifuge solutions at 4000 rpm for one hour | #*Centrifuge solutions at 4000 rpm for one hour | ||
#*Re-suspend particles in 140uL PBS (per solution) | #*Re-suspend particles in 140uL PBS (per solution) | ||
*Note: two samples of MHA/AuNPs were made: MHA dissolved in water & heated, MHA | *Note: two samples of MHA/AuNPs were made: MHA dissolved in water & heated, MHA in isopropanol & not heated | ||
===Fluorescence Data: 4 and 8 Repeat DNA with ThT=== | ===Fluorescence Data: 4 and 8 Repeat DNA with ThT=== |
Revision as of 12:38, 25 July 2013
Procedure Update, MHA Day One | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
July 25, 2013Procedure Update: MHA Silica-CoatingAccording to Aggregation and adhesion of gold nanoparticles in phosphate buffer saline, PBS and prolonged sonication actually cause aggregation in gold nanoparticle solutions: "results showed an optimized sonication time of 10 min to reach equilibrium for plain gold nanoparticles in water." Instead of using PBS to dilute all components, filtered MilliQ water will be used to dilute nanoparticles as well as MHA, as the organic solvent in solution prior to the actual silica-coating may have also been affecting observed the gold nanoparticle aggregation. Filtered MilliQ water will be brought up to room temperature as we've noticed cold solutions (whether water or PBS) have seemed to induce aggregation as well.
Fluorescence Data: 4 and 8 Repeat DNA with ThTFluorescence measurements taken and calibration curve constructed in order to show the increased fluorescence intensity with increases number of G-quadruplexes formed.<br.> <br.> <br.> <br.> <br.> <br.> <br.> |