pLV3-CW57-CMV-ssRFP-GFP-KDEL - 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 20μL sterile water to the plasmid dry powder to dissolve, and let stand at room temperature for 1 min; ② Mixing: Take 200μl competent cells and mix with 5-10μl plasmid DNA, and place on ice for 30 min; ③ Heat-shock introduction: Let stand at 42℃ for 90s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, and culture on a shaker at 37℃ for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100μl) of recovered cells onto LB plates with the corresponding antibiotic. Keep 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 plasmids according to experimental needs.
Reference Application	pLV3-CW57-CMV-ssRFP-GFP-KDEL is a lentiviral vector that uses the CMV promoter to drive the expression of the ssRFP-GFP-KDEL fusion reporter gene for monitoring endoplasmic reticulum autophagy (ER-phagy) activity.
Sharing Method	Public Welfare Sharing