BeNa Culture Collection
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| Culture medium | BNCC352243 |
| Description | LB nutrient agar |
| Composition | Yeast Extract:5.0g, Peptone:10.0g, NaCl:10.0g, Agar:15.0g, pH:7.0±0.2(25℃) |
| Growth conditions | 37 ℃; 18-24h; aerobic |
| Subculture procedure | 1. Place the competent cells in ice to melt (or let go of the heart or room temperature for a moment, quickly insert into the ice when the bacterial cells are in a mixed state of ice and water), add the target DNA (plasmid or ligation product) and gently mix by hand at the bottom of the EP tube, and let it stand on ice for 25 minutes. Heat shock in a 2.42 ℃ water bath for 45 seconds, quickly place it back on ice and let it stand for 2 minutes. Shaking will reduce the conversion efficiency. 3. Add 0.9 mL of LB medium to the centrifuge tube. Resuscitate at 4.37 ℃ and 225 rpm for 90 minutes. Centrifuge at 5.5000rpm for one minute to collect the bacteria. Take about 100 μ l of supernatant and gently blow and resuspend the bacterial blocks onto a culture medium containing the corresponding antibiotics. 6. Invert the plate and incubate overnight in a 37 ℃ incubator. If performing blue and white spot screening operation, incubate the plate at 37 ℃ for at least 17 hours. |
| Storage conditions | -80 ℃ |
| Sharing mode | Public welfare sharing |
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