IGEM:UC Berkeley/2006/Plasmid Transformation: Difference between revisions
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3. incubate on ice for 5 min. <br> | 3. incubate on ice for 5 min. <br> | ||
4. plate <br> | 4. plate <br> | ||
-JL_06/09/06 | -JL_06/09/06 <br> | ||
'''Transformation of Ligation mix''' | |||
1. Create cocktail on ice<br> | |||
*100 uL TGI cells<br> | |||
*15 uL KCM buffer<br> | |||
*25 uL water<br> | |||
*10 uL plasmid<br> | |||
2. Heat shock for 1.5 min at 42 C <br> | |||
3. Rescue with SOB media | |||
4. Incubate in 37 C shaker for 1 hour | |||
5. Plate | |||
-JL_06/14/06 |
Revision as of 11:09, 14 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.
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. Heat shock for 1.5 min at 42 C
3. Rescue with SOB media 4. Incubate in 37 C shaker for 1 hour 5. Plate
-JL_06/14/06