pEBB HA RelA - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	LB + Ampicillin, 37℃ (cloning strain DH5α, 7046bp)
Storage Conditions	2-8℃
Procedure	1. Dissolving: After receiving the plasmid dry powder, add 20μl sterile water to the bottom of the tube to dissolve the plasmid, and let it 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, and 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 the plates upright for 30min (ensure the agar surface is dry), then invert and culture for 12-16h until colonies appear. 7. Extraction: Pick single colonies into LB liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16h, and extract the plasmid as needed for the experiment.
Intended Use	Use: Protein expression, Host: Mammalian cells
Sharing Method	Public welfare sharing