pET-41a|394157 |BNCC

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pET-41a

Growth conditions	37°C; 18–24 hours; aerobic;
Subculture procedure	① Dissolution: Add 20 μL of sterile water to the plasmid Freeze dried pellet to dissolve it, and let stand at Room temperature for 1 minute; ② Mixing: Mix 200 μL of competent cells with 5–10 μL of plasmid DNA, and incubate on ice for 30 min; ③ Heat shock transformation: Incubate at 42°C for 90 s; ④ Shrink-wrap film well: Incubate on ice for 2 min; ⑤ Recovery culture: Add 800 μL of liquid medium to each tube and incubate on a shaking incubator at 37 °C for 1 h (150 r/min); ⑥ Screening: Spread an appropriate volume (100 μl) of resuspended cells onto the corresponding resistance LB agar plate; incubate the Agar plate upright for 30 min (ensuring the agar surface is completely dry), then incubate inverted for 12–16 h until colonies appear; ⑦ Isolation: Transfer a single colony to the corresponding LB liquid medium with the appropriate antibiotic resistance, incubate with shaking for 12–16 h, and extract the plasmid as required by the experiment.
Storage conditions	2–8°C
Sharing mode	Public welfare sharing