Difference between revisions of "Bacterial Transformation for Bio113"

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(Transforming DNA after GGA Ligation)
 
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# 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.  
 
# 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.
+
# Spread cells onto LB plates containing ampicillin.
 
You're done already!!
 
You're done already!!

Latest revision as of 17:49, 2 September 2022

Transforming DNA after GGA Ligation

  1. Thaw the competent cells on ice for 6 minutes. Each tube contains 50 µL of E. coli, JM109 cells.
  2. 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.
  3. Incubate on ice for 5 minutes.
  4. Spread cells onto LB plates containing ampicillin.

You're done already!!