pET-32a(+)-E-protein Plasmid|354678 |BNCC

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pET-32a(+)-E-protein plasmid

Subculture procedure	1. Dissolution: Upon receiving the plasmid Freeze dried pellet, add 20μl sterile water to the bottom of the tube to dissolve the plasmid. Allow to stand 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 pores: Ice bath for 2 minutes; 5. Repair culture: Add 800μl 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 LB liquid medium with the corresponding antibiotic, shake incubate for 12-16 hours, and extract the plasmid as required for the experiment.
Growth conditions	LB + Ampicillin, 37°C, Cloning Strain BL21(DE3)
Storage conditions	2-8°C
Safety level	1
Sharing mode	Public welfare sharing