BeNa Culture Collection
info@bncc.com
| Culture medium | BNCC361029 |
| Description | LB Medium with 50μg/mL Ampicillin |
| Composition | Yeast Extract:5.0g, Peptone:10.0g, NaCl:10.0g, pH:7.0(25℃) |
| Growth conditions | LB + antibiotics, 37°C |
| Subculture procedure | ①Dissolution: Add 20 μL sterile water to the plasmid powder for dissolution, let stand at Room temperature for 1 min; ② Mix: Add 200μl competent cells to 5-10μl plasmid DNA, mix thoroughly, and incubate on ice for 30 min; ③ Heat shock transformation: Incubate at 42°C for 90 s; ④ Shrink membrane wells: Ice bath for 2 min; ⑤ Repair culture: Add 800μl liquid medium to each tube, incubate on a 37°C shaking incubator for 1 h (150 rpm); ⑥ Screening culture: Spread an appropriate volume (100 μl) of thawed cells onto LB plates with the corresponding antibiotic. Incubate plates upright for 30 min (ensure agar surface is completely dry), then invert and incubate for 12-16 hours until colonies appear; ⑦ Extraction: Transfer a single colony to the corresponding antibiotic-containing LB liquid medium. Incubate with shaking for 12-16 hours. Extract the plasmid as required for the experiment. |
| Storage conditions | 2-8°C |
| Sharing mode | Public welfare sharing |
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