HKU-HKBU/29 July 2009

From 2009.igem.org

Co-transformation Transform two plasmids (A+, C+) to MG3.① Plac-CheZ-His-TT C+② Lac Iq A+

1 14:10 Subculture to warm room (37℃), shaking for 1.5h.
2 15:40 take the subculture bacteria out from the warm room.Freeze the bacteria in 0℃ ice water for
3 15min.Centrifuge in 4000rmp at 0℃ for 2min.Decent supernatant
4 15:40 Recover in warm room (37℃) for 1h.
5 18:40 take the bacteria out from warm room
6 19:05 Put plates to warm room (37℃) for overnight

14:10 Subculture to warm room (37℃), shaking for 1.5h.

15:40 take the subculture bacteria out from the warm room.Freeze the bacteria in 0℃ ice water for

15min.Centrifuge in 4000rmp at 0℃ for 2min.Decent supernatant

Add iced DI water, re-suspend Centrifuge in 8000rmp for 2min. Pipette away supernatant. (Above three steps should be repeated for 3 times.) Remove most supernatant, leaving behind~50μL. Electro-transformation

15:40 Recover in warm room (37℃) for 1h.

18:40 take the bacteria out from warm room

Transfer to agar plates with A+ & C+.