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HKU-HKBU/29 July 2009
From 2009.igem.org
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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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| - | + | {{Team:HKU-HKBU/style.css}} | |
| - | Transform two plasmids (A+, C+) to MG3. | + | {{Team:HKU-HKBU/script.js}} |
| - | ① Plac-CheZ-His-TT C+ | + | {{Team:HKU-HKBU/header}} |
| - | ② Lac Iq A+ | + | |
| + | 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.===== | ||
| - | =====15:40 take the subculture bacteria out from the warm room. | + | |
| - | Freeze the bacteria in 0℃ ice water for 15min. | + | =====15:40 take the subculture bacteria out from the warm room.Freeze the bacteria in 0℃ ice water for===== |
| - | Centrifuge in 4000rmp at 0℃ for 2min. | + | =====15min.Centrifuge in 4000rmp at 0℃ for 2min.Decent supernatant===== |
| - | 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===== | ||
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+
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+.
