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June 18, 2013
FCS
- Calibration:
- Alexa 532 Antibodies at 500pM and 100pM
- green fluorescent beads diluted 10000x
- Samples:
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
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