User:Melissa Novy/Notebook/CHEM-571/2012/11/04

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Objectives

  • Prepare 1 mL 0.4 M IPTG in autoclaved water.
  • Grow E. coli cells prepared on 2012/11/03 and induce protein expression of ADA.

Preparation of 0.4 M IPTG

  • Calculations:
  MW IPTG (β-D-1-thiogalactopyranoside): 238.31 g/mol
  0.4 M × 0.004 L = 0.0016 mol IPTG
  0.0016 mol IPTG × 238.31 g/mol IPTG = 0.3813 g IPTG
  Actual mass IPTG = 0.3813 g
  Actual concentration IPTG = 0.3813 g/0.004 L = 0.4 M
  • 1 mL of 0.4 M IPTG was added to each of four 1.5 mL microcentrifuge tubes to prepare for protein expression.

Protein Expression

  • Please refer to Keyun Wang's entry for protein expression procedure, absorbance data of cells to monitor cell growth, and analytic equipment settings.
  • Note that the cell pellets obtained after each centrifugation were resuspended in binding buffer made on 2012/09/26.