Difference between revisions of "Bacterial Transformation for Bio113"
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== Transforming DNA after GGA Ligation == | == Transforming DNA after GGA Ligation == | ||
| − | # Thaw the competent cells on ice for 6 minutes. Each tube contains 50 µL of cells. | + | # Thaw the competent cells on ice for 6 minutes. Each tube contains 50 µL of ''E. coli'', JM109 cells. |
# Add all 10 µL of your GGA ligation mixture to the thawed cells. Very gently mix the DNA and cells and return the cells to ice ASAP. | # Add all 10 µL of your GGA ligation mixture to the thawed cells. 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 13:06, 6 August 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 10 µL of your GGA ligation mixture to the thawed cells. 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!!
