pCT74-EGFP-GUS-HygR - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, pH: 7.0 (25°C)
Growth Conditions	37°C; 18-24h; Aerobic
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder to dissolve, and let stand at room temperature for 1 min; ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, and place on ice for 30 min; ③ Heat shock introduction: Let stand at 42°C for 90s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, and culture on a shaker at 37°C for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100 μl) of recovered cells on an LB plate with the corresponding antibiotic. Leave the plate upright for 30 min (make sure the agar surface is completely dry first), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single colonies into liquid Recommended Medium with the corresponding antibiotic in LB, culture with shaking for 12-16 h, and extract the plasmid as needed for the experiment.
Reference Use	pCT74-EGFP-GUS-HygR is an expression vector for filamentous fungi. It uses the ToxA promoter to drive the expression of EGFP (green fluorescent protein) and GUS (β-glucuronidase) genes, and carries the Hyg (hygromycin) resistance gene as a selection marker.
Sharing Method	Public Welfare Sharing