"

From 2010.igem.org

Laura's Lab Notebook:

Re-try failed transformation (failed 2X for Alex and Chris) Protocol followed:

1. get competent cells from -80oC freezer
2. set H2O bath to 42oC (already done)
3. add 50 uL cells to a 1.5 mL tube (keep tubes on ice)
4. add 50 ng or 2.5 uL circular DNA to E. Coli (variation: used 5.0 uL this time)
5. on ice 30 min
6. 24oC H2O bath 20 sec.
7. on ice 15 min
8. add 1 mL SOC, 2 hours at 37oC (rotating)
9. plate100uL on appropriate plate (kanamycin this time), incubate 12-16 hours before picking colonies
   1. variations: 200 uL plated this time, also plated on kan/X-gal/IPTG plate (plasmid inducible copy # by X-gal)