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Team:Stanford/NotebookPage/30 June 2010
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Karpadillo (Talk | contribs) (→Laura's Lab Notebook:) |
Karpadillo (Talk | contribs) (→Laura's Lab Notebook:) |
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#add 1 mL SOC, 2 hours at 37oC (rotating) | #add 1 mL SOC, 2 hours at 37oC (rotating) | ||
#plate100uL on appropriate plate (kanamycin this time), incubate 12-16 hours before picking colonies | #plate100uL on appropriate plate (kanamycin this time), incubate 12-16 hours before picking colonies | ||
| - | + | *variations: 200 uL plated this time, also plated on kan/X-gal/IPTG plate (plasmid inducible copy # by X-gal) | |
Latest revision as of 20:15, 19 July 2010
Laura's Lab Notebook:
Re-try failed transformation (failed 2X for Alex and Chris) Protocol followed:
- get competent cells from -80oC freezer
- set H2O bath to 42oC (already done)
- add 50 uL cells to a 1.5 mL tube (keep tubes on ice)
- add 50 ng or 2.5 uL circular DNA to E. Coli (variation: used 5.0 uL I0500 this time)
- on ice 30 min
- 24oC H2O bath 20 sec.
- on ice 15 min
- add 1 mL SOC, 2 hours at 37oC (rotating)
- plate100uL on appropriate plate (kanamycin this time), incubate 12-16 hours before picking colonies
- variations: 200 uL plated this time, also plated on kan/X-gal/IPTG plate (plasmid inducible copy # by X-gal)
