Minute Transformation of E. coli
1. Prewarm LB+antibiotic
plates at 37C.
2. Remove competent cells from –80C freezer and thaw on ice.
3. As soon as cells are thawed, immediately add 1-10ng DNA per 100ul
4. Incubate on ice for 5 minutes.
5. Spread cells on plates (100ul on one, 20-50ul on another).
6. Incubate at 37C overnight.Competent Cells
We currently have three types
of competent cells:
1. DH5? - general use
2. XL10-Gold – large or ligated DNA
3. XL1-Blue – unmethylated DNA; unlimited DNA or subcloning