pLV2-EF1a-hRluc-IRES-Hyg - Basic Information

Recommended Medium	Yeast extract: 5.0 g, Peptone: 10.0 g, NaCl: 10.0 g, pH: 7.0 (25°C)
Growth Conditions	37°C; 18-24 hours; Aerobic;
Storage Conditions	2-8°C
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: Let stand at 42°C for 90 s; ④ Membrane pore contraction: Place on ice for 2 min; ⑤ Recovery culture: Add 800 μL of liquid Recommended Medium to each tube, and culture on a shaker at 37°C for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μL) of recovered cells onto LB plates containing the appropriate antibiotic, keep the plates upright for 30 min (ensure the agar surface is completely dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single colonies into LB liquid Recommended Medium containing the appropriate antibiotic, culture with shaking for 12-16 h, and extract plasmid as needed for the experiment.
Reference Application	pLV2-EF1a-hRluc-IRES-Hyg is a lentiviral vector that uses the EF1a promoter to drive expression of the Renilla luciferase (hRluc) reporter gene, and is linked to the hygromycin resistance gene via an IRES element for selection of stable cell lines.
Sharing Method	Public welfare sharing

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