pLV3-CMV-MCS-3×Myc-Blast - 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 20 μL of sterile water to the lyophilized plasmid powder to dissolve it, and let it 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℃ 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 at 37℃ with shaking 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 (make sure the agar surface is completely 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, culture with shaking for 12-16 h, and extract the plasmid as needed for the experiment.
Reference Application	pLV3-CMV-MCS-3×Myc-Blast is a lentiviral expression vector that uses the strong CMV promoter to drive expression of the target gene from the multiple cloning site (MCS), allowing the target protein to be fused with a 3×Myc tag at the N-terminus or C-terminus for easy detection； It also carries a Blasticidin resistance gene for selection of stably transfected cell lines.
Sharing Method	Public welfare sharing