pET28a-STX1A(human) (1 Synonymous Mutation) - Basic Information

Recommended Medium	Yeast Extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	LB + Kanamycin, 37℃ (Cloning strain DH5α, 753bp)
Storage Conditions	2-8℃
Operating Steps	1. Dissolution: After receiving the plasmid dry powder, add 20μl sterile water to the bottom of the tube to dissolve the plasmid, and let stand at room temperature for 1min; 2. Mixing (plasmid adsorption): Mix 200μl competent cells + 5~10µl plasmid DNA, and place on ice for 30min; 3. Heat-shock introduction: Let stand at 42℃ for 90s; 4. Membrane pore contraction: Ice bath for 2min; 5. Recovery culture: Add 800µl LB liquid Recommended Medium to each tube, culture at 37℃ for 1h at 150 r/min; 6. Selection culture: Spread an appropriate volume (100 µl) of recovered cells onto LB plates with the corresponding antibiotic, keep plates upright for 30min (ensure the agar surface is dry), then invert and culture for 12-16h until colonies appear. 7. Extraction: Pick single monoclonal colonies into LB liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16h, and extract the plasmid according to experimental needs.
Reference Use	Vector host: E. coli, vector use: protein expression, gene species: human, gene type: ORF, prokaryotic resistance: Kan, fluorescent protein: unknown.
Sharing Method	Public welfare sharing

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