User:Elaine Marie Robbins/Notebook/Capstone/2012/07/12

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Fluorescence of Open MB Main project page
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Methods

  1. Make 3 solutions of MB [MB] = 10 nM:
    • MB alone
    • MB + complement, complement:MB = 10:1
    • MB + complement, complement:MB = 1:1
  2. Heat in heat block at 75°C for 30 min.
  3. Incubate at room temperature for 30 min.
  4. Run fluorescence.


  1. Make 4 solutions of MB [MB] = 200 nM:
    • MB alone, incubated at 75°C for 30 min.
    • MB alone, incubated at RT for 30 min.
    • MB + complement, complement:MB = 5:1, incubated at 75°C for 30 min.
    • MB + complement, complement:MB = 5:1, incubated at RT for 30 min.
  2. Incubate all solutions at room temp for 30 min.
  3. Run fluorescence.
  4. Take fluorescence of 100x diluted fluorescein.

Data

  • Fluorescence of MB open and closed, [MB] = 10 nM:
Image:12-07-12 fluorescence of MB 75C incubation.png
  • Fluorescence of MB open and closed, [MB] = 200 nM:
Image:12-07-12 fluorescence of MB 200 nM.png
Image:12-07-12 fluorescence of MB 200 nM with fluorescein.png

Fluorescein used as comparison.

  • Scan speed comparison:
Image:12-07-12 fluorescence of 200 nM MB scan speeds.png


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