User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/18: Difference between revisions
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==June 18, 2013== | ==June 18, 2013== | ||
===FCS=== | |||
*Calibration: | |||
**Alexa 532 Antibodies at 500pM and 100pM | |||
**green fluorescent beads diluted 10000x | |||
*Samples: | |||
**1-8 from 06/13/13 | |||
No fluorescence obvserved. | |||
===Sample Prep: Remake 727.5nM DNA/9.7nM AuNPs=== | |||
*Making 100mM DTT in 4% TEA | |||
*Extraction procedure with ethyl acetate | |||
*Combine equal volumes of 19.4nM AuNPs with 1455nM DNA from extraction to make 727.5nM DNA/9.4nM AuNP solution | |||
*Centrifuge four times, collecting four supernatants,at 4000rpm for 5 min intervals then re-dispersing in 50mM HEPES buffer | |||
*Take absorbance spectra for concentrated sample and four supernatants | |||
===Sample Prep: Remake 727.5nM DNA and 14.835nM AuNP Solutions=== | |||
*Take fluorescence spectra, exciting at 532nm for the Molecular Beacon data | |||
===Sample Prep: Additional DNA Concentrations for DNA/ThT Correlation Graph=== | |||
*Make the following solutions (use PBS for dilutions): | |||
**800nM DNA/2.5μM ThT | |||
**1091.25nM DNA/2.5μM ThT | |||
**1200nM DNA/2.5μM ThT | |||
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Revision as of 09:27, 18 June 2013
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June 18, 2013FCS
No fluorescence obvserved. Sample Prep: Remake 727.5nM DNA/9.7nM AuNPs
Sample Prep: Remake 727.5nM DNA and 14.835nM AuNP Solutions
Sample Prep: Additional DNA Concentrations for DNA/ThT Correlation Graph
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