pRE112 - Basic Information

Recommended Medium	Yeast extract: 5.0g, Peptone: 10.0g, NaCl: 10.0g, Agar: 15.0g, Chloramphenicol: 50μg/mL, pH: 7.0 (25°C)
Growth Conditions	LB + antibiotic, 37°C
Storage Conditions	2-8°C
Procedure	① Dissolution: Add 20μL sterile water to the plasmid dry powder for dissolution, and let stand at room temperature for 1 min; ② Mixing: Take 200μl competent cells and mix with 5-10μl plasmid DNA, and place on ice for 30 min; ③ Heat shock introduction: Let stand at 42°C for 90 s; 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 onto LB plates with the corresponding antibiotic resistance. Leave the plates upright for 30 min (the agar surface must be thoroughly dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into LB liquid Recommended Medium with the corresponding antibiotic resistance, culture with shaking for 12-16 h, and extract the plasmid as required by the experiment.
Sharing Method	Public welfare sharing