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Revision as of 22:14, 19 July 2015
Experiments & Protocols
Transformation protocol
- (If using part from the distribution: resuspend the DNA in 10 ul of MiliQ water, making sure that it turns red. Wait 10 minutes before adding the DNA to cells)
- Put a tube of NEB DH 5 alpha E. coli cells on ice and wait until they thaw completely. Divide the cells into 50 ul aliquotes.
- Add 1 ul of plasmid DNA to 50 ul of cells.
- Mix by carefully flicking the tube. Do not vortex or pipette in and out!
- Place the mixture on ice for 30 minutes.
- Heat shock the cells at 42 °C for 30 seconds and immediately put on back on ice.
- Keep cells on ice for next 5 minutes. Do not mix.
- Pipette 950 ul of SOC (or LB) media kept at room temperature into the mixture.
- Incubate the mixture at 37 °C for 60 minutes
- Plate 50 ul of cells to the plate with appropriate antibiotic and incubate overnight at 37 °C