Difference between revisions of "20.109(S18): Prep notes for M1"

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(M2D3)
(M2D5)
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==M2D5==
 
==M2D5==
 +
'''prior to laboratory:'''
 +
 +
*prepare 0.2 mM chymotrypsin in 1 mM HCl containing 2 mM CaCl<sub>2</sub>
 +
 
'''per team:'''
 
'''per team:'''
  
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*buffer reagents
 
*buffer reagents
 
**X &mu;L 1 M Tris-HCL pH=8
 
**X &mu;L 1 M Tris-HCL pH=8
**Y &mu;L 100 mM CaCl<sub>2</sub>
+
**Z &mu;L 0.2 mM chymotrypsin
**Z &mu;L chymotrypsin
+
  
 
==M2D6==
 
==M2D6==

Revision as of 21:54, 31 January 2018

20.109(S18): Laboratory Fundamentals of Biological Engineering

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Spring 2018 schedule        FYI        Assignments        Homework        Class data        Communication
       1. Assessing ligand binding        2. Measuring gene expression        3. Engineering biomaterials              


M1D1

  • restriction enzymes and buffers
  • filtered pipet tips

M1D2

prior to laboratory:

  • prepare 1% agarose gels, 1 per 2 teams
  • subculture BL21(DE3) pRSETb_FKBP12 1:10 into 50 mL LB containing 100μg / mL amp and 34 μg / mL cam, 1 per team

per team:

  • X μL 6x loading dye
  • Y μL 1 M IPTG

M2D3

prior to laboratory:

  • prepare dialysis buffer, X mL per team

per team:

  • lysis buffer reagents
  • 250 μL slurry in 2 mL microcentrifuge tube
  • 2.5 mL 1X PBS
  • Y μL 1 M MgCl2
  • 12 μL DNase
  • 2 mL 1X PBS containing 10 mM imidazole
  • 3.5 mL 1X PBS containing 250 mM imidazole

M2D4

prior to laboratory:

  • boil stained and unstained molecular weight standards, if required
  • prepare electrophoresis buffer, 1 chamber volume per 2 teams

per team:

  • 20 μL Laemmli buffer
  • 50 mL coomassie stain
  • BSA reagents
    • 110 μL albumin
    • 1300 μL 1X PBS
    • X μL BCA Reagent A
    • Y μL BCA Reagent B

M2D5

prior to laboratory:

  • prepare 0.2 mM chymotrypsin in 1 mM HCl containing 2 mM CaCl2

per team:

  • 18 μL 1 mg/mL FKBP12
  • buffer reagents
    • X μL 1 M Tris-HCL pH=8
    • Z μL 0.2 mM chymotrypsin

M2D6

per team:

  • 18 μL 1 mg/mL FKBP12

M2D7

per team:

  • none