"
Team:UC Davis/Notebook/Protocols
From 2012.igem.org
Protocols
•Add 20ul sterile H2O to desired well in distribution plate.
•Incubate at room temperature for ~10 minutes.
•Transfer resuspension to microcentrifuge tube.
Materials
•Competent Cells•DNA template
•800 ul LB
•LB+Antibiotic Plates
Procedure
•Thaw competent cells on ice.•Transfer 50 uL of competent cells to chilled falcon tubes.
•Add 1 uL of template to cells (2.5 uL if dilute).
•Incubate on ice for 30 minutes.
•Heat schock in 42 °C water bath for 90 seconds.
•Immediately place back onto ice for 2 minutes.
•Add 800 uL of LB to each tube.
•Incubate at 37 °C for 1 hour.
•Place 200 uL of the transformed cells on plates containing LB and the appropriate antibiotic.
•Incubate overnight at 37 °C.
