Protein Expression and Purification Series - Bio-Rad

CHAPTER 3B
ADVANCE PREP
INSTRUMENTATION PROCES
Protein Expression and Purification Series
Student Workstations
The following workstation setups are for the initial activities of 1) streaking a starter plate to produce single
bacterial colonies on LB/amp agar plates, 2) preparing overnight cell cultures, 3) subculturing and induction
of cells, and 4) preparing SDS-PAGE samples for later analysis.
1. Streak starter plates to produce single bacterial colonies on agar plates
Each student team requires the following items to streak a starter plate to produce single bacterial colonies:
Material Needed for Each Workstation Quantity
LB/amp plate 1
Sterile inoculating loop 1
2–20 µl adjustable-volume micropipet and tips 1
Marking pen 1
Common Workstation Quantity
Rehydrated BL21(DE3) E. coli containing pDHFR 1
Incubator set to 37°C 1
2. Overnight cell culture
Each student team requires the following items to prepare an overnight culture:
Material Needed for Each Workstation Quantity
LB/amp plate streaked with BL21(DE3) containing pDHFR 1
50 ml conical tube containing 15 ml sterile LB/amp broth 1
Sterile inoculating loop 1
20% sterile glucose 1 ml
100–1,000 µl adjustable-volume micropipet and tips 1
Marking pen 1
Common Workstation Quantity
Tube roller in a 37°C incubator, shaking water bath or shaking incubator set to 37°C 1
76 Chapter 3B: Advance Preparation for
Chromatography Instrumentation Protocols