Team:Purdue/E. coli transformation


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Transformation of E. Coli - Obtained materials – buckets of ice, 5 ml SOC, X21 Gold Competent Cells, Kan Plates, hot water bath, shaker in 37 degree room - Took XL-21 gold from -80, added 12μl to transformation tube and moved immediately to ice bucket - Added 1.5 μl luciferase plasmid DNA (with kanamycin resistance) to eppendorf tube - Prepared a control without the plasmid - Stored on ice for 30 minutes - Heat shock in 42 degree water bath for 28 sec - Moved to ice bucket for 2 minutes - Added 200μl SOC carefully to the control and the transformant - Moved the tubes to 37 degree shaker, set at 100 rpm, tilted at an angle for better mixing – for one hour - Obtained three kan plates and plated 50 μl control, 50 μl transform, and 100 μl of transform – streaked with glass beads - Stored plates in 37 degree room for 11 hours