User:Michael F. Nagle/Notebook/Chem 571/2012/09/19

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  1. Elution Buffer
  2. .02M Tris solution was needed for 10mL buffer.
    1. .02M Tris * .01L = .0002moles/liter
    2. .0002mol *121.14= .02423g Tris
    3. .249 was the actual amount weighed and put in 7mL water.
    4. HCl and NaOH were added dropwise until the pH was 7.53, between a target range of 7-8.
  3. .4g IPTG was weighed and placed in 4mL of buffer

1mL of IPTG solution was added to each expression culture.

  1. Buffer for binding was made with 20mM Tris, .5M NaOH and 20-40mM amidazole at a pH of 7.4
    1. .02M Tris solution *1L = .02moles Tris
    2. .02mol Tris * 121.14g = 2.4228g Tris
    3. .5M NaCl *1L =.5mol
    4. .5mol*58.439=29.2195gNaCl
    5. 29.2147gNaCl weighed
    6. 30mM imidazole *1L = .03mol imidazole
    7. .03mol*68.077g/mol=2.0423g imidazole
    8. 2.4721g Tris weighed
    9. HCl and NaOH were added dropwise until the pH was 7.4 as indicated by a pH meter.
  1. A buffer for elution was made with 20mM Tris, .5M NaCl and 500mM amidazole
    1. .02M Tris solution *.5L = .01moles Tris
    2. .01mol Tris * 121.14g = 1.2114g Tris
    3. 1.2164g Tris weighed
    4. .5M NaCl *.5 = .25mol
    5. .25mol*58.439g/mol=14.6098gNaCl
    6. .5M imidazole *.5L = .25mol imidazole
    7. .25mol*68.077g/mol=17.019g imidazole
  1. Tris buffer stock solution was inserted into HAuCl4 and BSA tubes at molarities of .05mM, .5mM, 5mM, 50mM, 100mM, 200mM, 500mM and 1M
    1. m1v1=m2v2
    2. 500mM*6mL=1000mM*x8##3mL
    3. 1000mM*6mL=1000mM*x
    4. 6mL
    5. 200mM*6mL=1000mM*x
    6. 1.2mL
    7. 100mM*6mL=1000mM*x
    8. .6mL
    9. 50mM*6mL=1000mM*x
    10. .3mL
    11. 5mM*6mL=1000mM*x
    12. .03mL
    13. .5mM*6mL=1000mM*x
    14. .003mL
    15. .05mM*6mL=1000mM*x
    16. .0003mL



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