User:Dhea Patel/Notebook/Phosphorylation/2012/07/24

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Objective

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Description

  1. Purified ATP was rotovapped with 10mL MeOH three times.
  2. 1.8mL DMF was added to the ATP under nitrogen. 0.301g DCC was added to the reaction mixture and allowed to react for 3 hours.
  3. 0.048805g (1,10)-Phenanthrolin-5-amine was dissolved in 20mL MeOH and added to the reaction mixture, along with 0.25mL TEA and allowed to react for 30 minutes.
  4. The reaction was quenched in 40mL H2O.
  5. The RuATP solution was run through the fast DEAE column, with the neon band running through with water. 0.5M TEAB was run through the column. The eluate solutions were covered and stored in the fridge over night.
  6. Fractions of the RuATP from the 11th of July continued to be rotovapped.
  7. TLC plates were run to determine which solvent would run the purified RuATP. During these tests, it was determined that 40% Acetonitrile solution with some sodium nitrate would run purified ATP.

Calculations

0.25mmol (1,10)-Phenantolin-5-amine

0.25mmol ÷ 1000mmol/mol × 195.22g/mol = 0.048805g

Data

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Notes

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