Team:Purdue/E. coli transformation
From 2010.igem.org
Materials
- buckets of ice
- 5 ml SOC
- X21 Gold Competent Cells
- Kan Plates
- hot water bath
- shaker in 37 degree room
Procedure
- 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