User:Elaine Marie Robbins/Notebook/CHEM-496/2011/11/08

From OpenWetWare
Jump to navigationJump to search
Purification <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Objective

To purify the proteins expressed in cells frozen on 09/28/11.

Description

  1. Make solutions for tomorrow.
    • 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)
  2. Filter both 1 L solutions.
  3. Defrost cells from 09/28/11 in a water bath.
  4. Sonicate cells: 30 s on, 30 s off 3 times.
  5. Centrifuge at 17000 rpm for 1.5 hr.
  6. Run dialysis overnight on supernatant in the 2 batches of 4L of 50 mM Tris (pH 9).

Data

Notes