pX459 V2.0|390539 |BNCC

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pX459 V2.0

Growth conditions	LB + Ampicillin, 37°C (Cloning strain: Stbl3, Origin: U6, Repository: pUC)
Subculture procedure	①Dissolution: Add 20μL sterile water to the plasmid Freeze dried pellet and dissolve; let stand at Room temperature for 1 min; ②Mixing: Take 200μl competent cells and mix with 5-10μl plasmid DNA; place on ice for 30 min; ③Heat shock transformation: Incubate at 42°C for 90 sec; ④ Cooling: Place on ice for 2 min; ⑤ Growth: Add 800 μL liquid medium to each tube and incubate at 37°C on a shaker (150 rpm) for 1 h; ⑥ Screening culture: Agar plate an appropriate volume (100 μl) of thawed cells onto LB agar 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-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