pCAMBIA3300-35S-MCS-DsRed-35S-Bar - 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 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: Place 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 incubate at 37°C with shaking at 150 r/min for 1 h; ⑥ Selection culture: Spread an appropriate volume (100 μL) of recovered cells onto LB plates with the appropriate antibiotic, leave the plates upright for 30 min (ensure the agar surface is thoroughly dry), then invert and incubate for 12–16 h until colonies appear; ⑦ Extraction: Pick single colonies into liquid Recommended Medium with the appropriate antibiotic, culture with shaking for 12–16 h, and extract the plasmid as required for the experiment.
Intended Use	pCAMBIA3300-35S-MCS-DsRed-35S-Bar is a binary expression vector for plants. It uses dual CaMV 35S promoters to drive expression of the target gene in the multiple cloning site and the red fluorescent protein DsRed, respectively, and carries the Bar gene (herbicide resistance) as a plant selection marker and kanamycin resistance as a prokaryotic resistance marker.
Sharing Method	Public welfare sharing

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