pCMV-N-Flag-BiolD2 - 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 h; Aerobic
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20 μL of sterile water to the plasmid lyophilized powder to dissolve, and let stand at room temperature for 1 min; ② Mixing: Mix 200 μL of competent cells with 5-10 μL of plasmid DNA, and place on ice for 30 min; ③ Heat-shock introduction: Let stand at 42°C for 90 s; ④ Membrane pore contraction: Place on ice for 2 min; ⑤ Recovery culture: Add 800 μL of 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 recovered cells onto an LB plate with the corresponding antibiotic. Leave the plate upright for 30 min (make sure the agar surface is thoroughly 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 as needed for the experiment.
Reference Application	pCMV-N-Flag-BioID2 is a proximity protein biotin labeling plasmid for mammalian cells. It uses the CMV promoter to efficiently express a fusion protein of a bait protein with a 3X Flag tag and BioID2. By biotinylating proximal proteins within approximately 10 nm of the target protein in living cells, it is used to study protein-protein interactions.
Sharing Method	Public welfare sharing