pADV-EF1a-MCS - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25℃)
Growth Conditions	37℃； 18-24 hours； Aerobic；
Storage Conditions	2-8℃
Procedure	① Dissolution: Add 20 μL of sterile water to the lyophilized plasmid 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℃ 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 culture on a shaker at 37℃ 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. Leave the plates upright for 30 min (the agar surface must be completely dry), then invert and culture for 12-16 h until colonies appear； ⑦ Extraction: Pick single monoclonal colonies and inoculate them 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	pADV-EF1a-MCS is an adenoviral expression vector that uses the EF1a promoter to drive high-level expression of the target gene in the multiple cloning site (MCS) in mammalian cells. It is suitable for recombinant adenovirus packaging and protein overexpression studies.
Sharing Method	Public welfare sharing