"
Team:Paris Saclay/Protocols/Transformation of supercompetent Ecoli cells with CaCl2
From 2014.igem.org
(Difference between revisions)
(Created page with "= '''Transformation of E. coli cells with CaCl2''' = E.coli is not naturally competent and there are several ways to prepare competent cells for plasmid DNA transformation. This...") |
(→Transformation of E. coli cells with CaCl2) |
||
| Line 8: | Line 8: | ||
Day 1: Incubate one colony from LB plate in LB liquid medium. Shake at 37 °C overnight. | Day 1: Incubate one colony from LB plate in LB liquid medium. Shake at 37 °C overnight. | ||
| - | Day 2: Dilute the overnight cell culture 100 to 200 times in 100 ml LB medium. Shake vigorously at 37 °C. | + | Day 2: Dilute the overnight cell culture 100 to 200 times in 100 ml LB medium. Shake vigorously at 37 °C until OD650nm between 0,2 and 0,3. |
| + | |||
| + | Place in an ice bath for few minutes and centrifuge the cells for 10 min at 4,000 rpm at 4 °C. | ||
Revision as of 10:34, 12 July 2014
Transformation of E. coli cells with CaCl2
E.coli is not naturally competent and there are several ways to prepare competent cells for plasmid DNA transformation. This is the chemical method with CaCl2.
1.Preparation of competent E.coli cells
Day 1: Incubate one colony from LB plate in LB liquid medium. Shake at 37 °C overnight.
Day 2: Dilute the overnight cell culture 100 to 200 times in 100 ml LB medium. Shake vigorously at 37 °C until OD650nm between 0,2 and 0,3.
Place in an ice bath for few minutes and centrifuge the cells for 10 min at 4,000 rpm at 4 °C.
