pLV3-CMV-EGFP-MCS-3×HA-EF1a-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 20 μL of sterile water to the plasmid dry powder to dissolve, and let stand at room temperature for 1 min; ② Mixing: Mix 200 μL of competent cells with 5-10 μL of plasmid DNA, and place on ice for 30 min; ③ Heat shock introduction: Place at 42℃ for 90 s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800 μL of liquid Recommended Medium to each tube, and culture on a shaker at 37℃ for 1 h (150 r/min); ⑥ Screening culture: Spread an appropriate volume (100 μL) of recovered cells onto LB plates with the corresponding antibiotics. Leave the plates upright for 30 min (the agar surface must be dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single colonies into liquid Recommended Medium with the corresponding antibiotics, culture with shaking for 12-16 h, and extract the plasmid as needed for the experiment.
Intended Use	pLV3-CMV-EGFP-MCS-3×HA-EF1a-Puro is a lentiviral vector that uses the CMV promoter to drive the expression of EGFP fused with the target gene in the multiple cloning site (MCS) and a 3×HA tag. A separate EF1a promoter drives Puromycin resistance for selection of stable cell lines.
Sharing Method	Public welfare sharing