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Team:University of Lethbridge/Notebook/GeneralLabAugst
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| + | Tranformation of pUC19 | ||
| + | |||
| + | 1. Thaw E.coli DH5a cells on ice. | ||
| + | 2. Add pUC19 DNA, pipette gently to mix (1μl of plasmid). | ||
| + | 3. Let sit for 30 minutes on ice. | ||
| + | 4. Incubate cells for 30 seconds at 42oC. | ||
| + | 5. Incubate cells on ice for 2 min. | ||
| + | 6. Add 1 mL SOC at room temp. | ||
| + | 7. Incubate for 1 hour at 37oC on shaker. | ||
| + | 8. Spread 100-300 μl onto a plate made with appropriate antibiotic. | ||
| + | 9. Grow overnight at 37 °C. | ||
Revision as of 23:07, 3 August 2008
August 03, 2008
Nathan Phillips
Tranformation of pUC19
1. Thaw E.coli DH5a cells on ice. 2. Add pUC19 DNA, pipette gently to mix (1μl of plasmid). 3. Let sit for 30 minutes on ice. 4. Incubate cells for 30 seconds at 42oC. 5. Incubate cells on ice for 2 min. 6. Add 1 mL SOC at room temp. 7. Incubate for 1 hour at 37oC on shaker. 8. Spread 100-300 μl onto a plate made with appropriate antibiotic. 9. Grow overnight at 37 °C.
