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From 2013.igem.org

1.Preparation for the competent cells in the -80’c refrigerator, and target DNA molecular.Then refresh the competent cells in the 0℃ icy water.
3.Mix the two components and put the EP tube in the icy water for half an hour.
3.Next put them in the 42℃ water for 90 secs.
4.Inject the LB solution into the EP tube and culture them at least 40 mins.
5.Paste them in the plate.