pCRE-miniP-Fluc-hPEST-Neo-Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25°C)
Growth Conditions	37°C; 18-24 hours; Aerobic;
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder for dissolution, let stand at room temperature for 1 min; ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, place on ice for 30 min; ③ Heat shock: Place at 42°C for 90 s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, culture at 37°C with shaking 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 plates upright for 30 min (ensure the agar surface is completely dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into LB liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract the plasmid as needed for the experiment.
Reference Application	pCRE-miniP-Fluc-hPEST-Neo is a CRE pathway luciferase reporter plasmid for mammalian cells. It utilizes the CRE promoter to drive expression of the unstable Fluc-hPEST reporter gene and carries neomycin (Neo/G418) resistance for stable selection.
Sharing Method	Public Welfare Sharing