Difference between revisions of "E. coli transformation"
| Line 1: | Line 1: | ||
| + | Using the puch tool, puch out the appropriate DNA. Between puches | ||
| + | |||
Add 2µL of DNA in TE to 50µL of competent cells (TOP10) | Add 2µL of DNA in TE to 50µL of competent cells (TOP10) | ||
| − | Heat shock at 42ºC for 60 sec. | + | Allow the DNA and competent cells to sit on ice for 20 minutes |
| + | |||
| + | Heat shock at 42ºC for 60 sec in water bath. | ||
Recover on ice for 5 min. | Recover on ice for 5 min. | ||
| Line 7: | Line 11: | ||
Add 200 µL SOC media | Add 200 µL SOC media | ||
| − | Incubate at 37ºC for 2 hr. | + | Incubate at 37ºC for 2 hr while the tubes are rotating. |
Revision as of 19:54, 20 March 2008
Using the puch tool, puch out the appropriate DNA. Between puches
Add 2µL of DNA in TE to 50µL of competent cells (TOP10)
Allow the DNA and competent cells to sit on ice for 20 minutes
Heat shock at 42ºC for 60 sec in water bath.
Recover on ice for 5 min.
Add 200 µL SOC media
Incubate at 37ºC for 2 hr while the tubes are rotating.