User:Elaine Marie Robbins/Notebook/Capstone/2012/06/26

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Methods

  1. Make 8 solutions:
    • 3 of MB alone:
      1. 260 μL tris buffer.
      2. Incubate at room temp for 30 min.
      3. Add 6.11 μL/1.22 μL of MB or 12.2 μL 100x diluted MB.
      4. Incubate at 37°C for 60 min.
    • 3 of MB and cells:
      1. 250 μL diluted cell extract containing 100,000 cells and 10 μL tris buffer.
      2. Incubate at room temp for 30 min.
      3. Add 6.11 μL/1.22 μL of MB or 12.2 μL 100x diluted MB.
      4. Incubate at 37°C for 60 min.
    • AuNP/TS primers:
      1. 3 μL AuNP/TS primers, 3.5μL tris buffer, 1 μL dNTP, 2.5 μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells.
      2. Incubate at room temp for 30 min.
      3. Add 1.22 μL of MB.
      4. Incubate at 37°C for 60 min.
    • Primers only:
      1. 6.5 μL AuNP/TS primers, 1 μL dNTP, 2.5 μL PCR buffer, and 250 μL diluted cell extract containing 100,000 cells.
      2. Incubate at room temp for 30 min.
      3. Add 1.22 μL of MB.
      4. Incubate at 37°C for 60 min.
  2. Make a 50 µM solution of fluorescein in 100 mM sodium borate buffer, pH 9.5.

Data

12-06-26 fluorescence of MB and cells.png

This contradicts data from 06/25/12, when the MB and cells had a higher fluorescence intensity than solutions containing AuNP/primer and dNTPs.