pCDNA3.1(+) - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	LB + antibiotic, 37℃
Storage Conditions	2-8℃
Procedure	① Dissolving: Add 20μL sterile water to the plasmid dry powder and let stand at room temperature for 1 min; ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA and place on ice for 30 min; ③ Heat-shock transformation: Place at 42℃ for 90s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube and culture at 37℃ with shaking (150 r/min) for 1h; ⑥ Selection culture: Spread an appropriate volume (100 μl) of recovered cells onto LB plates with the corresponding antibiotic. Leave plates upright for 30 min (the agar surface must be dry), then invert and culture for 12-16h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with the corresponding antibiotic and culture with shaking for 12-16h. Extract the plasmid as needed for the experiment.
Reference Use	pCDNA3.1(+) is a classic and highly efficient constitutive mammalian cell expression vector. It uses the cytomegalovirus immediate-early promoter to achieve high-level, sustained expression of target genes in a variety of mammalian cell lines. The vector backbone has been optimized for stable replication in both mammalian cells and E. coli, making it one of the preferred tools for gene function research, protein expression and production, and stable cell line construction.
Sharing Method	Public welfare sharing