pMIGR1 - 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 Dissolve: Add 20 μL sterile water to the plasmid dry powder to dissolve it, and let stand at room temperature for 1 min; ② Mix: Take 200 μL competent cells and mix with 5–10 μL plasmid DNA, then place on ice for 30 min; ③ Heat-shock introduction: Place at 42°C for 90 s; ④ Contract membrane pores: Place on ice for 2 min; ⑤ Recovery culture: Add 800 μL liquid Recommended Medium to each tube, and culture at 37°C on a shaker for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μL) of the 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.
Intended Use	pMIGR1 is a retroviral empty vector (deposited by Addgene, catalog #27490), constructed based on the MSCV (murine stem cell virus) backbone. It uses the MESV promoter to drive gene expression and co-expresses EGFP green fluorescent protein through an IRES element, and is used for efficient gene overexpression and fluorescence tracing in mammalian cells, such as T cells.
Sharing Method	Public-interest sharing