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(New page: =====Co-transformation Transform two plasmids (A+, C+) to MG3. ① Plac-CheZ-His-TT C+ ② Lac Iq A+===== =====14:10 Subculture to warm room (37℃), shaking for 1.5h.===== =====15:40...)
 
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=====Co-transformation  
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Transform two plasmids (A+, C+) to MG3.
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① Plac-CheZ-His-TT  C+
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② Lac Iq  A+=====
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Co-transformation Transform two plasmids (A+, C+) to MG3.① Plac-CheZ-His-TT  C+② Lac Iq  A+
=====14:10 Subculture to warm room (37℃), shaking for 1.5h.=====
=====14:10 Subculture to warm room (37℃), shaking for 1.5h.=====
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=====15:40 take the subculture bacteria out from the warm room.
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Freeze the bacteria in 0℃ ice water for 15min.
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=====15:40 take the subculture bacteria out from the warm room.Freeze the bacteria in 0℃ ice water for=====
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Centrifuge in 4000rmp at 0℃ for 2min.
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=====15min.Centrifuge in 4000rmp at 0℃ for 2min.Decent supernatant=====
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Decent supernatant=====
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Add iced DI water, re-suspend
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Centrifuge in 8000rmp for 2min.
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Pipette away supernatant.
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(Above three steps should be repeated for 3 times.)
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Remove most supernatant, leaving behind~50μL.
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Electro-transformation
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=====15:40 Recover in warm room (37℃) for 1h.=====
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=====18:40 take the bacteria out from warm room=====
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Transfer to agar plates with A+ & C+.
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=====19:05 Put plates to warm room (37℃) for overnight=====

Latest revision as of 14:32, 16 October 2009

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

Contents

[hide]
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+.

19:05 Put plates to warm room (37℃) for overnight