PiggyBac-CMV-MCS-EF1a-puro - Basic Information

Recommended Medium	Yeast Extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25°C)
Growth Conditions	37°C; 18-24h; Aerobic
Storage Conditions	2-8°C
Procedure	①Dissolution: Add 20μL sterile water to the plasmid dry powder to dissolve, and let stand at room temperature for 1 min; ②Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, and place on ice for 30 min; ③Heat-shock introduction: Let stand at 42°C for 90 s; ④Pore contraction: Place on ice for 2 min; ⑤Recovery culture: Add 800μl liquid Recommended Medium to each tube, and culture in a 37°C shaker for 1 h (150 r/min); ⑥Screening culture: Spread an appropriate volume (100 μl) of recovered cells on LB plates with the corresponding antibiotic. Keep the plates upright for 30 min (make sure the agar surface is completely dry), then invert and culture for 12-16 h until colonies appear; ⑦Extraction: Pick single monoclonal colonies into LB liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract the plasmid as required by the experiment.
Reference Application	PiggyBac-CMV-MCS-EF1a-Puro is a eukaryotic expression vector based on the PiggyBac transposon system, using the strong CMV promoter to drive efficient expression of the target gene in the multiple cloning site (MCS); At the same time, the EF1a promoter independently drives expression of the Puromycin resistance gene for screening stably integrated cell lines.
Sharing Method	Public Welfare Sharing