pBlueScript II SK(+) - 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, 2961bp)
Storage Conditions	2-8℃
Procedure	① Dissolving: 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℃ 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 at 37℃ on a shaker for 1 h (150 r/min); ⑥ Selection culture: Spread an appropriate volume (100μL) of recovered cells onto LB plates with the corresponding antibiotic, leave the plates upright for 30 min (the agar surface must be dry), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick a single monoclonal colony into liquid Recommended Medium with the corresponding antibiotic, culture with shaking for 12-16 h, and extract the plasmid according to experimental needs.
Sharing Method	Public welfare sharing