User:Elaine Marie Robbins/Notebook/Capstone/2012/06/27

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Methods

  1. Make 2 solutions of telomerase:
    • Primers only (no AuNP):
      1. Mix 6.5 μL 100x diluted primers, 1 μL dNTP, 2.5 μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells.
    • AuNP/TS primers:
      1. Mix 3 μL AuNP/TS primers, 1 μL dNTP, 2.5 μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells.
  2. Make a solution containing 260 μL tris buffer.
  3. Make a solution containing 250 μL diluted cell extract containing 100,000 cells, and 10 μL tris buffer.
  4. Incubate all solutions at room temperature for 30 min.
  5. Add 1.22 μL MB to all solutions.
  6. Incubate all solutions at 37oC for 1 hr.
  7. Take fluorescence data.
  8. Take fluorescence of dilutions of fluorescein solution.

Data

  • Fluorescence of MB and cells:
Image:12-06-27 fluorescence of MB and cells.png


  • Fluorescence emission spectra for fluorescein dilutions, excited at 470 nm:
Image:12-06-27 fluorescein spectra.png
Image:12-06-27 fluorescein calibration curve.png


  • Charts of relative fluorescence intensity:
Image:12-06-27 relative fluorescence intensity chart number of cells.png


Image:12-06-27 relative fluorescence intensity type of synthesis.png


  • Peak fluorescence vs MB concentration for components of telomerase solution mixed with MB:
Image:12-06-27 calibration curve for telomerase solution components.png


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