IGEM:UC Berkeley/2006/Plasmid Transformation

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KCM-competent cell heat shock (don't use this procedure for ligation products or cotransformations)

1. create cocktail on ice
*10 uL KCM buffer
*70 uL water
2. heat shock for 1.5 min at 42 C
3. incubate on ice for 5 min.
4. plate


Transformation of Ligation mix( use for ligation product transformations or cotransformations)

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


Electro Shock Competence

1. Set volts: 1.60 (slow thaw cells on ice) Use 2YT media or SOB media
2. Put 50 uL of cells into clean eppendorf and limit bubbles. Add 1 uL of ligation mixture and stir to mix.
4. Put into holds, well seated in back
5. After beep, rescue with 500 uL(have ready) of media. Transfer all volume back to eppendorf, no longer on ice.



  1. 0.5M KCl
  2. 0.15M CaCl2
  3. 0.25M MgCl2
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