pNFAT-pTA-GFP-Neo - 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 lyophilized plasmid powder to dissolve it, and let it 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: Ice bath for 2 min; ⑤ Recovery culture: Add 800 μL of liquid Recommended Medium to each tube, and incubate on a shaker at 37°C 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 resistance. Leave the plates upright for 30 min (be sure the agar surface is dry first), then invert and incubate for 12-16 h until colonies appear; ⑦ Extraction: Pick a single colony into liquid Recommended Medium with the corresponding antibiotic resistance, culture with shaking for 12-16 h, and extract the plasmid as needed for the experiment.
Reference Application	pNFAT-pTA-GFP-Neo is an NFAT transcription factor reporter plasmid used in mammalian cells. It uses a pTA minimal promoter to drive expression of GFP green fluorescent protein and carries a neomycin (Neo/G418) resistance gene. It is commonly used in T cell activation, calcium signaling pathway, and related drug screening studies.
Sharing Method	Public welfare sharing

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