Difference between revisions of "Bacterial Transformation for Bio113"
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# Thaw the competent cells on ice for 6 minutes. Each tube contains 50 µL of ''E. coli'', JM109 cells. | # Thaw the competent cells on ice for 6 minutes. Each tube contains 50 µL of ''E. coli'', JM109 cells. | ||
| − | # Add all | + | # Add all 50 µL of your ''E. coli'' cells to the GGA ligation mixture. Very gently mix the DNA and cells and return the cells to ice ASAP. |
# Incubate on ice for 5 minutes. | # Incubate on ice for 5 minutes. | ||
# Add 60 µL SOC media with no antibiotic to your transformed cells. Spread cells onto LB plates containing ampicillin. | # Add 60 µL SOC media with no antibiotic to your transformed cells. Spread cells onto LB plates containing ampicillin. | ||
You're done already!! | You're done already!! | ||
Revision as of 19:21, 23 October 2018
Transforming DNA after GGA Ligation
- Thaw the competent cells on ice for 6 minutes. Each tube contains 50 µL of E. coli, JM109 cells.
- Add all 50 µL of your E. coli cells to the GGA ligation mixture. Very gently mix the DNA and cells and return the cells to ice ASAP.
- Incubate on ice for 5 minutes.
- Add 60 µL SOC media with no antibiotic to your transformed cells. Spread cells onto LB plates containing ampicillin.
You're done already!!
