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

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(Autocreate 2012/11/04 Entry for User:Melissa_Novy/Notebook/CHEM-571)
Current revision (16:08, 10 November 2012) (view source)
 
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==Entry title==
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==Objectives==
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* Insert content here...
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* Prepare 1 mL 0.4 M IPTG in autoclaved water.
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* Grow <i>E. coli</i> cells prepared on [[User:Melissa_Novy/Notebook/CHEM-571/2012/11/03|2012/11/03]] and induce protein expression of ADA.
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** There were no deviations from the [[AU_Biomaterials_Design_Lab:Protocols/Protein_Expression|protein expression]] protocol.
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==Preparation of 0.4 M IPTG==
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* Calculations:
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  '''MW IPTG (β-D-1-thiogalactopyranoside): 238.31 g/mol'''
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  '''0.4 M × 0.004 L = 0.0016 mol IPTG'''
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  '''0.0016 mol IPTG × 238.31 g/mol IPTG = 0.3813 g IPTG'''
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  '''Actual mass IPTG = 0.3813 g'''
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  '''Actual concentration IPTG = 0.3813 g/0.004 L = 0.4 M'''
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* 1 mL of 0.4 M IPTG was added to each of four 1.5 mL microcentrifuge tubes to prepare for protein expression.
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==Protein Expression==
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* Please refer to [[User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I/2012/11/04|Keyun Wang's entry]] for protein expression procedure, absorbance data of cells to monitor cell growth, and analytic equipment settings.
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* Note that the cell pellets obtained after each centrifugation were resuspended in binding buffer made on [[User:Melissa_Novy/Notebook/CHEM-571/2012/09/26|2012/09/26]].
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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.



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