User:Andrew K Farag/Notebook/CHEM-571/2014/02/12

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Protocol

  1. Synthesize new NCs (3 sets)
  2. Data Analysis
  3. Continue precipitation and collection of QDs


General Protocol

  1. In test tube mix 180 uL of 5mM Zinc Nitrate and 20 uL of 5mM Manganese Acetate
  2. Add 200 uL of 40 mM Ammonium Acetate
  3. Add 200 uL of Protein:ligand in various ratios from 0% to 100% protein
  4. Adjust pH to 8.2 using roughly 250 uL of 10mM Ammonium Hydroxide
  5. Incubate at room temp for 1 hour
  6. Add 200 uL of 5mM Sodium Sulfide drop wise, then cover the test tubes
  7. A VWR Analog Vortex Mixer was used to vortex each test tube for 30 seconds
  8. Incubate for five-six days at 37 degrees Celsius
  • Also need to create stock solutions for BSA, 1-thioglycerol, and Sodium Sulfide

BSA-L-cysteine NC table of volumes

% BSA 5 mM Zinc Nitrate (uL) 5mM Manganese Acetate (uL) 41.65 mM Ammonium Acetate (uL) 11uM BSA* (uL) 10uM L-Cysteine (uL) 10mM Ammonium Hydroxide (uL) 5 mM Sodium Sulfide* (uL) Total Volume (mL)
80 540 60 600 480 120 3000 600 5.4
85 540 60 600 510 90 3000 600 5.4
90 540 60 600 540 60 3000 600 5.4
95 540 60 600 570 30 3000 600 5.4
100 540 60 600 600 0 3000 600 5.4

BSA-GSH NC Table of volumes

% BSA 5 mM Zinc Nitrate (uL) 5mM Manganese Acetate (uL) 41.65 mM Ammonium Acetate (uL) 10uM BSA* (uL) 10uM GSH (uL) 10mM Ammonium Hydroxide (uL) 5 mM Sodium Sulfide* (uL) Total Volume (mL)
70 540 60 600 420 180 3000 600 5.4
75 540 60 600 450 150 3000 600 5.4
80 540 60 600 480 120 3000 600 5.4
85 540 60 600 510 90 3000 600 5.4
90 540 60 600 540 60 3000 600 5.4
95 540 60 600 570 30 3000 600 5.4
100 540 60 600 600 0 3000 600 5.4

BSA-1-thioglycerol table of volumes

  • This is the first round of synthesis, therefore a ratio at which the NC's are stable in solution has not been identified and we will conduct standard ratios.
% BSA 5 mM Zinc Nitrate (uL) 5mM Manganese Acetate (uL) 41.65 mM Ammonium Acetate (uL) 11uM BSA* (uL) 10uM 1-Thioglycerol (uL) 10mM Ammonium Hydroxide (uL) 5 mM Sodium Sulfide* (uL) Total Volume (mL)
0 540 60 600 0 600 3000 600 5.4
10 540 60 600 60 540 3000 600 5.4
20 540 60 600 120 480 3000 600 5.4
30 540 60 600 180 420 3000 600 5.4
40 540 60 600 240 360 3000 600 5.4
50 540 60 600 300 300 3000 600 5.4
60 540 60 600 360 240 3000 600 5.4
70 540 60 600 420 180 3000 600 5.4
80 540 60 600 480 120 3000 600 5.4
90 540 60 600 540 60 3000 600 5.4
100 540 60 600 600 0 3000 600 5.4