Stbl3 Chemotroph Competent Cells|365123 |BNCC

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Stbl3 Chemically Competent Cell

Growth conditions	37°C; 18-24h; aerobic
Subculture procedure	1. Thaw competent cells on ice (or briefly place in palm or at room temperature until cells are in a slushy state, then quickly return to ice). Add target DNA (plasmid or ligation product) and gently mix by tapping the bottom of the Eppendorf tube. Incubate on ice for 25 minutes. 2. Heat shock at 42°C for 45 seconds, then immediately return to ice and let stand for 2 minutes. Agitation reduces transformation efficiency. 3. Add 700μl antibiotic-free sterile LB medium to the centrifuge tube. Mix thoroughly and incubate at 37°C, 200 rpm for 60 minutes. 4. Centrifuge at 5000 rpm for 1 minute to collect cells. Retain approximately 100 μl supernatant, gently resuspend the pellet by pipetting, and plate onto LB medium containing the appropriate antibiotic. 5. Place plates upside down in a 37°C incubator overnight. For blue-white screening, incubate plates at 37°C for at least 17 hours.
Storage conditions	-80°C
Sharing mode	Public welfare sharing