Team:ULaVerne Collab/notebook
PROTOCOLS
Transformation Protocol (E. coli)
1. Pre-chill microcentrifuge tube and thaw competent cells on ice.
2. Add 50uL of competent cells into the chilled microfuge tube.
3. Add 4uL of DNA into the microcentrifuge tube.
4. Incubate on ice on 30 minutes.
5. Heat shock at 42 degrees Celsius for 45 sec.
6. Incubate cells on ice for 2 minutes.
7. Add 200uL of SOC media into the microcentrifuge tube.
8.Incubate and shake at 37 degree Celsius for 1-2 hours.
9. Pipette 200uL of cells into LB plate with ampicillin resistance.
10. Incubate plate overnight at 37 degrees Celsius.
2. Add 50uL of competent cells into the chilled microfuge tube.
3. Add 4uL of DNA into the microcentrifuge tube.
4. Incubate on ice on 30 minutes.
5. Heat shock at 42 degrees Celsius for 45 sec.
6. Incubate cells on ice for 2 minutes.
7. Add 200uL of SOC media into the microcentrifuge tube.
8.Incubate and shake at 37 degree Celsius for 1-2 hours.
9. Pipette 200uL of cells into LB plate with ampicillin resistance.
10. Incubate plate overnight at 37 degrees Celsius.
Transformation Protocol (E. coli)
1. Pre-chill microcentrifuge tube and thaw competent cells on ice.
2. Add 50uL of competent cells into the chilled microfuge tube.
3. Add 4uL of DNA into the microcentrifuge tube.
4. Incubate on ice on 30 minutes.
5. Heat shock at 42 degrees Celsius for 45 sec.
6. Incubate cells on ice for 2 minutes.
7. Add 200uL of SOC media into the microcentrifuge tube.
8.Incubate and shake at 37 degree Celsius for 1-2 hours.
9. Pipette 200uL of cells into LB plate with ampicillin resistance.
10. Incubate plate overnight at 37 degrees Celsius.
2. Add 50uL of competent cells into the chilled microfuge tube.
3. Add 4uL of DNA into the microcentrifuge tube.
4. Incubate on ice on 30 minutes.
5. Heat shock at 42 degrees Celsius for 45 sec.
6. Incubate cells on ice for 2 minutes.
7. Add 200uL of SOC media into the microcentrifuge tube.
8.Incubate and shake at 37 degree Celsius for 1-2 hours.
9. Pipette 200uL of cells into LB plate with ampicillin resistance.
10. Incubate plate overnight at 37 degrees Celsius.
