User:Eleni N. Kalivas/Notebook/CHEM-571/2014/02/12

From OpenWetWare

Jump to: navigation, search
Project name Main project page
Previous entry      Next entry

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)
805406060048012030006005.4
85540606005109030006005.4
90540606005406030006005.4
95540606005703030006005.4
10054060600600030006005.4

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)
705406060042018030006005.4
755406060045015030006005.4
805406060048012030006005.4
85540606005109030006005.4
90540606005406030006005.4
95540606005703030006005.4
10054060600600030006005.4

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)
054060600060030006005.4
10540606006054030006005.4
205406060012048030006005.4
305406060018042030006005.4
405406060024036030006005.4
505406060030030030006005.4
605406060036024030006005.4
705406060042018030006005.4
805406060048012030006005.4
90540606005406030006005.4
10054060600600030006005.4


Personal tools