User:Paul Rothenberg/Notebook/Pauls Notebook/2013/03/18

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18 March 2013

  • Objectives:
  • 1. Run UV-VIS and Fluorescence spectroscopy on prepared solutions of proteins in DMSO
  • Procedure:
  • 1. A control sample of 200 µL DMSO was run in UV-VIS and Fluorescence spectroscopes
  • 2. 200 µL of Mb, DMSO, Crown, and Phosphate solution was run
  • 3. 190 µL of DMSO and 10 µL of Mb, DMSO, Crown, Phosphate was run
  • 4. 199 µL of DMSO and 1 µL of Mb, DMSO, Crown, Phosphate was run
  • 5. 25 µL of [4] was diluted with 175 µL of DMSO

  • 6. 200 µL of Mb, Acetate, DMSO, Crown was run
  • 7. 20 µL of [6] was diluted with 180 µL DMSO (10x factor)
  • 8. 20 µL of [7] was diluted with 180 µL DMSO (10x factor)
  • Data:
  • 1. The control spectra showed signs of contamination in the cuvettes; aquaregia was used to clean the cuvettes
  • 2. DMSO showed a peak between 200-300 nm in UV-VIS spectrum and a large peak in Fluorescence spectroscopy
  • 3. The 200 µL of DMSO, Crown, Phosphate was too concentrated − diluted
  • 4. [3] was still too concentrated under fluorescence
  • 5. [4] was too concentrated − removed and saved; 25 µL of [4] was added along with 175 µL DMSO
  • 6. Concentration of [5] did not max out the fluorimeter

  • 7. [6] was highly concentrated under UV-VIS, and not run under Fluorescence
  • 8. [7] was too concentrated under fluorescence, and diluted
  • 9. [8] did not max out the fluorimeter or the UV-VIS



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