Jonesma Lab:Protocols:Ecolitransformation

E. coli transformation

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1. Take aliquot of E. coli (Top10) from -80°C storage
2. Allow to thaw on ice
3. Add DNA (1μL of a vector or 10μL of a ligation reaction)
4. Chill mixture on ice for 30 min
5. Turn on waterbath to 42°C
6. Move bacteria to waterbath (at 42°C) for 30s
7. Immediately put back on ice for 5 min to chill
8. Add 1mL of LB
9. Shake at 37°C for 1 hr
10. Spin at 3000rpm for 15 min
11. Take off most of excess liquid and resuspend in approx 100μL LB
12. Plate on LB plates containing your antibiotic of choice O/N at 37°C