Bacterial Transformation for Bio113
From GcatWiki
Revision as of 19:20, 18 July 2017 by Macampbell (talk | contribs) (Created page with " == Transforming DNA after GGA Ligation == # Thaw the competent cells on ice for 6 minutes. Each tube contains 50 µL of cells. You can use these straight. Diluting up to 10...")
Transforming DNA after GGA Ligation
- Thaw the competent cells on ice for 6 minutes. Each tube contains 50 µL of cells. You can use these straight. Diluting up to 10 fold will work for ligations as well as direct transformations of minipreps.
2) Very gently, aliquot cells into smaller volumes (we have used as low as 20 µL of cells with 5 µL of ligation) using pre-chilled microfuge tubes.
3) Add 1 - 5 µL of ligation mixture (can go as high as 10 µL for larger volumes of cells).
4) Incubate on ice for 5 minutes.
5) Add SOC media with no antibiotic to a final volume of 60 - 100 µL. Spread cells onto plates containing ampicillin*. You must let this sit for 30 minutes in the liquid at 37 C before plating if the antibiotic is not ampicillin.
You're done already!!
