PACYC184|353817 |BNCC

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pACYC184

Growth conditions	LB+chloramphenicol, 37 ℃ (cloned strain DH5a)
Subculture procedure	1. Dissolution: After receiving the plasmid powder, please add 20 μ l of sterile water to the bottom of the tube, dissolve the plasmid, and let it stand at Room temperature for 1 minute; 2. Mixing (adsorption plasmid): Mix 200 μ l of competent cells and 5-10 µ l of plasmid DNA, and place on ice for 30 minutes; 3. Heat shock introduction: let stand at 42 ℃ for 90 seconds; 4. Shrinkage film pores: ice bath for 2 minutes; 5. Repair culture: Add 800 µ l LB liquid medium to each tube and incubate at 37 ℃ for 1 hour at 150 r/min; 6. Screening and cultivation: Apply an appropriate volume (100 µ l) of revived cells onto corresponding resistant LB plates, place them upright in the Agar plate for 30 minutes (the agar surface must be dry), invert and culture for 12-16 hours, and colonies will appear. 7. Extraction: Select monoclonal colonies into the corresponding resistant LB liquid medium, shake and culture for 12-16 hours, and extract plasmids according to experimental needs.
Storage conditions	2-8 ℃
Sharing mode	Public welfare sharing