User:Dhea Patel/Notebook/Phosphorylation/2012/06/22

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  1. The mixture was filtered under the hood. Clear crystals were collected.
  2. The filtered solution was run in the fast-flowing DEAE column (which had run 100mL H2O immediately before). A small orange band formed at the top of the column.
  3. After the sample was added, 125mL of H2O was run.
  4. This was followed by 60mL 0.1M TEAB. This gradient continued until the column cleared. (Gradient: H2O, 0.1M TEAB, 0.2 M TEAB, 0.3 M TEAB, 0.4 M TEAB, 0.5 M TEAB, 0.6 M TEAB, 1 M TEAB).
  5. 200mL of 1M TEAB was run
  6. 150mL of 1 M TEAB : 1 M NaCl
  7. 300mL of H2O (to clean out the column)


Mixture after filtering: opaque solution resembling orange juice.

After the mixture was added to the column, the liquid that was leaving the column as the sample was going into the column material was cloudy white. After adding more water to the column the run-through was cloudy until the water added to the column was clear. The run-throughs remained clear through the TEAB gradient. An orange band did form in the column and began to move slowly with 0.2 M TEAB.


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