pCDF1-MCS2-EF1-Puro - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	37℃； 18-24 hours； Aerobic；
Storage Conditions	2-8℃
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder to dissolve it, 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 introduction: Let stand at 42℃ for 90 s; ④ Shrinking membrane pores: Ice bath for 2 min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, and culture on a shaker at 37℃ for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μl) of recovered cells on LB plates with the corresponding antibiotic, leave the plates upright for 30 min (the agar surface must be thoroughly dry first), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid LB Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract plasmids according to experimental needs.
Intended Use	pCDF1-MCS2-EF1-Puro is a lentiviral expression vector based on the FIV (feline immunodeficiency virus) backbone. It uses the CMV promoter to drive high-level expression of the target gene in the multiple cloning site (MCS2), and the EF1α promoter independently drives the Puromycin resistance gene for selection of stably transduced cell lines.
Sharing Method	Public welfare sharing