User:Laura Flynn/Notebook/Experimental Biological Chemistry/2011/11/08

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Objective

To purify the proteins expressed in cells frozen on 09/29/11.

Procedure

Solutions were made for the experiment being performed on 11/09/11.

  • 1 L 50 mM Tris (pH 9.02)
  • 1 L 50 mM Tris + 1 M NaCl (pH 9.05)
  • 2 batches of 4 L 50 mM Tris (pH 9.00)

Both of the previous 1 Liter solutions were then filtered.

The cells from 09/28/11 were defrosted using a warm water bath. The cells were sonicated in cycles of 30 seconds on, 30 seconds off three times. The cells were centrifuged at 17000 rpm for 1.5 hours. The centrifuged material was discarded and dialysis was run overnight on the supernatant in 50 mM Tris (pH 9)

Results

Notes