IGEM:American University/2009/Notebook/Advanced Experimental Chemistry: Fun Times in Beeghly/2013/04/03

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Welcome to Advanced Experimental Chemistry

Today's Procedure

Reaction the Hydrogels with Fluorescent Dye:

  1. dye in dmso, add a little and mix with film in water
  2. Let dye, ascorbic, cuso4 and hydrogel solution over night to react with dye
  3. Cu(I) is unstable so add CuSO4 (Cu(II)) and ascorbic acid
  4. after 24 hours take a uv-vis of the hydrogel soaked in a known volume of dH20

Dye Calculatations

  • 17.24µL butyne-1-ol in the hydrogel
  • (17.24/70.09)*.926= 2.08E-4 moles Butyne-1-ol
  • rxn is 1:1 however we want 1.5 * excess of dye
  • Dye= azide-flour 488: mw 563.35g/mol
  • 563.25g/mol * 2.28E-4moes = 1.28g * 1.5 =0.193 g
    • not enough dye so we cut off a portion of the hydrogel to use and dye

Mass of cut hydrogel

  • mass of whole= 12.1419 g
  • mass of cut= 0.8810
  • percent of whole = 7.26%
  • Original amount of dye needed= 0.014g
    • however, we only have 0.005 g

Copper sulfat concentration

  • 249.8*0.001moles/l*0.01l= 0.0025 g of copper sulfate
  • MADE A STOCK SOLUTION OF .025g/ml so 100µl =0.0025g of copper sulfate

L-Ascorbic acid

  • 176.13g/mol * .1mol/L * 0.0L= 0.1761g
  • made stock solution of 1.761g per 10ml so 1 ml = .1761g

range of dye volumes tested since did not have enough made a stock solution of 5 mg of dye in 1 ml of dmso

  • ranges: 500, 200, 100, 10 and 1 µL of dye
  • Added to hydrogels 9 ml of water + dye + 100µL of copper sulfate + 1 ml of ascorbic acid