Kai Yuet/Protocols:Transformation

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=Bacterial Transformation (KPY)=

Materials

 * Chemically Competent E. Coli Cells
 * 10 μL Ligation Mixture

Procedures

 * 1) Retrieve chemically competent cells from -80°C freezer.
 * 2) Thaw cells for 5 minutes on ice.
 * 3) Add 2.5 μL of 10 μL ligation mixture into the microcentrifuge tubes containing the cells, stir gently, and then agitate the tubes.
 * 4) Ice the cells for 30 minutes.
 * 5) Heat shock the cells for 5 minutes at 37°C in the water bath.
 * 6) Incubate the cells on ice for 2 minutes.
 * 7) Add 500 μL of LB (no antibiotic) to the cells.
 * 8) Place the cells back into water bath for another 30 minutes of incubation.
 * 9) Spread the mixture of LB and cells onto a plate with the appropriate antibiotic.
 * 10) Grow overnight at 37°C.