pMV261-mcherry-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 20μL sterile water to the plasmid dry powder for dissolution, let stand at room temperature for 1 min; ② Mixing: Mix 200μl competent cells with 5-10μl plasmid DNA, place on ice for 30 min; ③ Heat shock introduction: Let stand at 42℃ for 90s; ④ Membrane pore contraction: Ice bath for 2 min; ⑤ Recovery culture: Add 800μl liquid Recommended Medium to each tube, culture at 37℃ on a shaker 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 plates upright for 30 min (ensure the agar surface is thoroughly dry first), then invert and culture for 12-16 h until colonies appear; ⑦ Extraction: Pick single monoclonal colonies into liquid Recommended Medium with the corresponding antibiotic resistance, culture with shaking for 12-16 h, and extract plasmids as needed for the experiment.
Reference Use	pMV261-mCherry-MCS is a shuttle expression vector for mycobacteria, utilizing the Mycobacterium smegmatis hsp60 promoter to drive the expression of the red fluorescent protein mCherry. It contains a multiple cloning site (MCS) for convenient insertion of target genes and carries kanamycin resistance.
Sharing Method	Public welfare sharing