Cell Transformation was performed
- BL21(DE3) cells thawed gently on ice
- The competent cells were mixed. 100 μL cells were added to three separate 14-ml BD Falcon polypropylene round bottom tubes.
- The three tubes were incubated on ice for 10 minutes, swirling gently ever 2 minutes
- 5 μL, 10 μL, and 15 μL of ligated DNA were added to one of the three tubes, and swirled gently
- The three tubes were incubated on ice for 30 minutes
- Each of the three tubes was heat-pulsed for 45 seconds in a 42°C water bath
- The three tubes were incubated on ice for 2 minutes
- 0.9 ml of LB medium was added to each of the three tubes and the tubes were incubated at 37 °C for 1 hour while shaking at 225 rpm.
- 200 μL of cells were plated and spread with a sterile spreader
- The cells were incubated overnight at 37°C
Cell transformation was unsuccessful