User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/28

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June 28, 2013

Notes from Meeting

  • Find averages and standard deviation of inconsistent DNA/ThT spectra for calibration curve
    • Fit Gaussian for inconsistent spectra (use Igor Pro program)
  • Check literature for AuNP@SiO2 shift and silica-shell radius calculations
  • Label axes for FCS data
  • FCS spectra for fluorescent beads in prezi
  • FCS: increase time as concentration decreases
  • Repeat old AuNP procedure with solutions from new procedure

FCS Measurements

  • Prepare Samples
    • 10000x diluted green fluorescent beads
    • 100, 80, 60, 20pM Oligo D
    • 10, 1, 5nM, 500, 100, 50, 25pM DNA/MB samples

Results and Notes

Calibration graphs:<br.> FCS data 2013 0628 calibration graphs.PNG<br.> <br.>

  • Oligo D samples were run for 300s, the 60pM and 80pM samples are still inconsistent with the trend of the other concentrations
  • The first round of MB/DNA samples were run for 300s, results had lots of noise and inconsistencies
  • The second round of MB/DNA samples were run for 700s to eliminate noise, only the 25pM sample lied outside the trend of the other concentrations
  • Future work:
    • Always align laser with 10000x diluted green fluorescent beads, include spectra for comparison
    • Create more samples for MB/DNA with 100pM intervals from 1nM to 100pM, and a 75pM sample
    • Create more samples for Oligo D at 50, 30, and 10pM concentrations
    • Make samples by serial dilutions after hybridization (?)
    • For pM concentrations increase time to 700s to eliminate noise from spectra
      • Increase time even further for concentrations lower than 25pM (?)
    • Attach AuNPs in proper ratio at successfully run concentrations of MB/DNA