Help:Transformation Protocol
Revision as of 18:42, 10 June 2009 by Vinoo (Talk | contribs) (New page: We have found that the following protocol ensures a high efficiency for transforming competent cells. This may be particularly useful if you are finding that your transformations are not w...)
We have found that the following protocol ensures a high efficiency for transforming competent cells. This may be particularly useful if you are finding that your transformations are not working, or yielding few colonies.
- Allow the cells to thaw
- Transfer 50ul into a 1.5 or 2ml centrifuge tube. Transform the 50ul of cells with 1ul of DNA
- Incubate on ice for 30 minutes
- Heat shock for 1 minute at 42C
- Hold on ice for 5 minutes
- Add 200 μl of SOC
- Incubate at 37 C for 2 hours on rotor
- Plate 20 μl and 200ul of the transformation on the appropriate antibiotic plates, and spread using sterile 3.5 mm glass beads