pLV3-EF1a-EGFP-Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	37℃； 18-24h； Aerobic
Storage Conditions	2-8℃
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder to dissolve, and let stand at room temperature for 1min； ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, and place on ice for 30min； ③ Heat shock transformation: Let stand at 42℃ for 90s； ④ Membrane pore contraction: Ice bath for 2min； ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, and culture on a shaker at 37℃ for 1h（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 30min（the agar surface must be dry first）, then invert and culture for 12-16h until colonies appear； ⑦ Extraction: Pick single monoclonal colonies into LB liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16h, and extract the plasmid as required by the experiment
Reference Application	pLV3-EF1a-EGFP is a lentiviral expression vector that uses the EF1a promoter to drive constitutive expression of EGFP (enhanced green fluorescent protein), for labeling transfected/infected cells or as a fluorescent tracing reporter gene. It usually carries a Puromycin resistance gene for selection of stable cell lines.
Sharing Method	Public Welfare Sharing