Protein Expression and Purification Series - Bio-Rad

Protein Expression and Purification Series
Measure OD600 of subculture and induce GST-DHFR-His expression
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After one hour of incubation measure the OD600 of your subculture.
Blank a spectrophotometer at 600 nm with 1 ml LB/amp broth.
Add 1 ml of subculture to a semi-microvolume cuvette and measure the OD600 of the undiluted
subculture and record below.
OD600 of subculture = ____________________________
Add 25 µl of 1 M IPTG to the subculture in the 500 ml Erlenmeyer flask and continue incubating and
shaking the culture at 37ºC for 4–24 hours. During this time period, most of the resources of the E. coli
will go toward producing the recombinant GST-DHFR-His protein rather than cell division, though cell
division does still occur.
Record the time that you start and end your induction below.
Induction Start Time: _______________________________
Induction End Time: ________________________________
Chapter 5: 100 ml Culture Protocol for
Chromatography Instrumentation Purification
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CHAPTER 5
100 ml CULTURE
PROTOCOL