pREDKI|354472 |BNCC

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pREDKI

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 + Ampicillin, 30°C (Cloning strain DH5α, 9028 bp)
Subculture procedure	1. Dissolution: Upon receiving the plasmid powder, add 20μl sterile water to the bottom of the tube to dissolve the plasmid. Incubate at Room temperature for 1 minute. 2. Mixing (Plasmid Adsorption): Mix 200μl competent cells + 5~10μl plasmid DNA thoroughly. Incubate on ice for 30 minutes. 3. Heat shock transformation: Incubate at 42°C for 90 seconds; 4. Shrink membrane wells: Ice-bathe for 2 minutes; 5. Repair culture: Add 800 µl LB liquid medium to each tube; incubate at 37°C for 1 hour at 150 rpm; 6. Screening culture: Spread an appropriate volume (100 µl) of the revived cells onto LB plates with the corresponding antibiotic. Incubate plates upright for 30 min (ensure agar surface is dry), then invert and incubate for 12–16 hours until colonies appear. 7. Extraction: Transfer a single colony to the corresponding antibiotic-supplemented 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