pLV3-CMV-MCS-3×HA-VC155-Puro - 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 for dissolution 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: Ice bath for 2 min; ⑤ Recovery culture: Add 800 μL of liquid Recommended Medium to each tube and culture at 37°C on a shaker for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μL) of recovered cells onto LB plates with the corresponding antibiotic. Leave the plates upright for 30 min (the agar surface must be thoroughly dry), then invert and culture for 12–16 h until colonies appear; ⑦ Extraction: Pick single colonies into liquid Recommended Medium with the corresponding antibiotic and culture with shaking for 12–16 h. Extract the plasmid as needed for the experiment.
Intended Use	pLV3-CMV-MCS-3×HA-VC155-Puro is a lentiviral expression vector that uses the CMV promoter to drive fusion expression of the gene of interest in the multiple cloning site (MCS) with VC155 (the C-terminal fragment of a fluorescent complementation protein), carries a 3×HA tag for convenient detection, and enables selection of stable cell lines through puromycin resistance. It is suitable for bimolecular fluorescence complementation (BiFC) experiments for protein–protein interactions.
Sharing Method	Public welfare sharing