pLV3-CMV-mKate2-Blast - Basic Information

Recommended Medium	Yeast extract: 5.0 g, Peptone: 10.0 g, NaCl: 10.0 g, 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 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℃ 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℃ 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 dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract the plasmid according to experimental needs.
Reference Use	pLV3-CMV-mKate2-Blast is a lentiviral expression vector that uses the CMV promoter to drive expression of the far-red fluorescent protein mKate2, and carries a Blasticidin resistance gene for selection of stable cell lines.
Sharing Method	Public welfare sharing