IGEM:UC Berkeley/2006/Plasmid Transformation: Difference between revisions
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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. | 3.5 If antibiotic marker is anything but Amp then rescue cells in 50uL LB for one hour at 37C.<br> | ||
4. plate <br> | 4. plate <br> | ||
-JL_06/09/06 <br> | -JL_06/09/06 <br> | ||
Revision as of 16:54, 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
