IGEM:UC Berkeley/2006/Plasmid Transformation

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Line 7: Line 7:
  2. heat shock for 1.5 min at 42 C <br>
  2. heat shock for 1.5 min at 42 C <br>
  3. incubate on ice for 5 min. <br>
  3. incubate on ice for 5 min. <br>
 +
3.5 If antibiotic marker is anything but Amp then rescue cells in 50uL LB for one hour at 37C.
  4. plate <br>
  4. plate <br>
-JL_06/09/06 <br>
-JL_06/09/06 <br>

Revision as of 18:53, 19 June 2006

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

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