IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/07/01/Zhangs

From OpenWetWare
Jump to: navigation, search

(1) Got 3×100μL DH5α competent cells from the -70℃ refrigerator and divided them equally into six EP tubes, each for 50μL.
(2) Wait for the solution to melt.
(3) Add 10μL ligation system: 1-23L 7:1, 1-23L 3:1, 1-4H 7:1, 1-4H 3:1, 1-23L Control and 1-4H Control.
(4) Keep in ice for 30min.
(5) Heat shock in 42℃ for 90s.
(6) Put the competent cells back into the ice box immediately for 2min.
(7) Add 200μL antibiotic LB in a super clean bench and recover for 45min.
(8) Centrifuge ,discard 100μL supernatant and smear the remaining 100μL onto the Amp+ plate.
(9) Culture under 37℃.