pMSGV-MCS-P2A-EGFP - 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 lyophilized plasmid powder to dissolve it, and let it stand at room temperature for 1 min; ② Mixing: Take 200 μL of competent cells and mix with 5-10 μL of plasmid DNA, then place on ice for 30 min; ③ Heat-shock introduction: Place 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 on a shaker at 37°C 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 according to experimental needs.
Intended Use	pMSGV-MCS-P2A-EGFP is a retroviral expression vector that uses the MSCV LTR promoter to drive expression of the target gene inserted into the multiple cloning site (MCS), and achieves independent co-expression with EGFP green fluorescent protein through the P2A self-cleaving peptide, enabling long-term stable expression and fluorescence tracing of the target gene.
Sharing Method	Public Welfare Sharing