IGEM:UC Berkeley/2006/Plasmid Transformation

KCM-competent cell heat shock

1. create cocktail on ice

  *20 uL TGI cells

  *10 uL KCM buffer

  *70 uL water

  *1 uL plasmid

2. heat shock for 1.5 min at 42 C 

3. incubate on ice for 5 min. 

3.5 If antibiotic marker is anything but Amp then rescue cells in 50uL LB for one hour at 37C.

4. plate 

-JL_06/09/06

Transformation of Ligation mix

1. Create cocktail on ice

  *100 uL TGI cells

  *15 uL KCM buffer

  *25 uL water

  *10 uL plasmid

2. Incubate on ice for 10 min.

3. Heat shock for 1.5 min at 42 C 

4. Rescue: add 150uL of SOB media 

5. Incubate in 37 C shaker for 1 hour

6. Plate

-JL_06/14/06