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HKU-HKBU/29 July 2009
From 2009.igem.org
(Difference between revisions)
| Line 8: | Line 8: | ||
=====15:40 take the subculture bacteria out from the warm room.Freeze the bacteria in 0℃ ice water for===== | =====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===== | =====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===== | ||
Revision as of 14:31, 16 October 2009
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 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+.
