User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/07/12

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July 12, 2013

Notes from Meeting

  • Adjust range and use SymphoTime to better fit FCS graphs in Igor
  • Use error bars on calibration graphs (for Igor <N> values)
  • Focus on attaching AuNPs to DNA & hybridizing MB to run on FCS
  • Use a range of volumes of TES in MHA silica-coating procedure to change silica-coating thickness
    • Take absorbance data to see differences between solution, then use TEM imaging to verify presence of silica-coating
  • Presentation at the end of 10 weeks

DNA and AuNP Attachment Procedure


  • Done by Allison Alix.<br.>
  • Extraction Procedure:
    • Combine 250uL 4.45uM DNA (diluted in PBS) with 250uL 100nM DTT in 4% TEA
    • React for 10 minutes
    • Extract DTT with 4 2mL aliquots of ethyl acetate (DTT will partition into ethyl acetate, DNA will partition into aqueous solution)
  • Combine 231.3uL 2.226uM DNA, 231.3uL 29.67nM 14.7nm AuNPs, and 37.3uL sodium citrate buffer prepared 07/11/2013 <br.>
    • Final Concentrations in Solution:
      • DNA: 1.03uM
      • AuNP: 13.73nM
      • Sodium citrate: 20mM
  • React for 10 minutes
  • Centrifuge 4 times, extracting the supernatant each time and then redispersing in 250uL 50nM HEPES buffer
  • Take absorbance of final solution and three supernatants

Absorbance Data