User:Michael F. Nagle/Notebook/Chem 571/2012/09/19: Difference between revisions
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==Entry title== | ==Entry title== | ||
* | #.02M Tris solution was needed for 10mL buffer. | ||
## .02M Tris * .01L = .0002moles/liter | |||
##.0002mol *121.14= .02423g Tris | |||
##.249 was the actual amount weighed and put in 7mL water. | |||
## HCl and NaOH were added dropwise until the pH was 7.53, between a target range of 7-8. | |||
#.4g IPTG was weighed and placed in 4mL of buffer | |||
1mL of IPTG solution was added to each expression culture. | |||
#Buffer for binding was made with 20mM Tris, .5M NaOH and 20-40mM amidazole at a pH of 7.4 | |||
##.02M Tris solution *1L = .02moles Tris | |||
##.02mol Tris * 121.14g = 2.4228g Tris | |||
##.5M NaCl *1L =.5mol | |||
##.5mol*58.439=29.2195gNaCl | |||
##29.2147gNaCl weighed | |||
##30mM imidazole *1L = .03mol imidazole | |||
##.03mol*68.077g/mol=2.0423g imidazole | |||
##2.4721g Tris weighed | |||
##HCl and NaOH were added dropwise until the pH was 7.4 as indicated by a pH meter. | |||
#A buffer for elution was made with 20mM Tris, .5M NaCl and 500mM amidazole | |||
##.02M Tris solution *.5L = .01moles Tris | |||
##.01mol Tris * 121.14g = 1.2114g Tris | |||
##1.2164g Tris weighed | |||
##.5M NaCl *.5 = .25mol | |||
##.25mol*58.439g/mol=14.6098gNaCl | |||
##.5M imidazole *.5L = .25mol imidazole | |||
##.25mol*68.077g/mol=17.019g imidazole | |||
Revision as of 10:37, 19 September 2012
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Entry title
1mL of IPTG solution was added to each expression culture.
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