User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/18

Replication Attempts

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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 Absorbance and Fluorescence Data Absorbance data taken for DNA/ThT samples, (needs to be rerun due to inconsistencies)and extracted AuNPs with DNA and four supernatants Samples might be remade before re-running Fluorescence data taken for DNA/ThT samples, DNA and ThT comparison