pCMV-HA-MCS-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 20μL sterile water to the plasmid dry powder to dissolve, let stand at room temperature for 1min； ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, place on ice for 30min； ③ Heat shock introduction: Let stand at 42℃ for 90s； ④ Membrane pore contraction: Ice bath for 2min； ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, culture at 37℃ with shaking for 1h（150 r/min）； ⑥ Selection culture: Spread an appropriate volume（100 μl）of recovered cells onto LB plates with corresponding resistance, leave plates upright for 30min（agar surface must be dry），then invert and culture for 12-16h until colonies appear； ⑦ Extraction: Pick single colonies into LB liquid Recommended Medium with corresponding resistance, culture with shaking for 12-16h, extract plasmid according to experimental needs
Reference Application	pCMV-HA-MCS is a plasmid vector used for transient expression in mammalian cells, driven by the strong CMV promoter. It carries an HA tag at the N-terminus for easy detection of the target protein, and contains a multiple cloning site（MCS）for gene cloning.
Sharing Method	Public Welfare Sharing