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Team:Brown/Protocols/Transformation
From 2010.igem.org
Bacterial Cell Transformation (Adrian 7/8/10)
- To transform competent cells. 200 µl of the suspension in a precooled microfuge tube (add cold 50mM CaCl2 to bring volume to 200ul). Add DNA mixture to the cells (20-30pg), mixing gently with your pipette on ice. Cover the tube and incubate for 20-30 min. on ice.
- Prepare a beaker of water (tap water is fine) at 42oC. Heat shock the cells by placing the glass culture tube at 42oC for 1 min., and then back on ice for 2 min. Add 0.1 µl LB media and then spread the cells over one or two LV/Amp plates, invert in 37oC incubator for overnight.
For blue/white screening of pGEM:
- Add 50uL of X-gal (from 20mg/mL stock)
- Add 2uL of IPTG (IM stock)
