User:Catherine Koenigsknecht/Notebook/Experimental Biological Chemistry/2011/09/14: Difference between revisions
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(Autocreate 2011/09/14 Entry for User:Catherine_Koenigsknecht/Notebook/Experimental_Biological_Chemistry) |
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==Description== | ==Description== | ||
# | # The E.coli cultures grown overnight were centrifuged at 4500rpm for 15 minutes at 4°C. | ||
# The supernatent was discarded and the cell pellets were each resuspended in 1mL of rich broth (4mL total). | |||
# The resuspended pellets were evenly divided into four 2800mL flasks that contained 1L of rich broth with 100μg/mL ampicillin. | |||
# These flasks were then shook at 165rpm at 37°C for about three hours. At this point 1mL of 0.1M IPTG was added to each flask. | |||
# The cells continued to grow and now express protein for about three more hours at 165rpm at 37°C. | |||
# The cultures were centrifuged at 4500rpm for 15 minutes at 4°C. In total the four pellets were resuspended in 200mL of column buffer. The cultures were then stored at -20°C. | |||
*Column Buffer: 20mM Tris-HCl (pH 7.4), 200mM NaCl, and 1mM EDTA | |||
==Data== | ==Data== |
Revision as of 08:16, 20 September 2011
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