Instructions Ⅰ. Plasmid dry powder (stored at minus 20 degrees, shelf life 90 days, transportation at room temperature) 1. After receiving the dry powder plasmid, please centrifuge at 5000rpm for 1min, and then add 20μl of sterile water to dissolve the plasmid; 2. Thaw the corresponding competent on an ice box, take 2 μl of plasmid and add it to 100 μl of competent for 30min; 3. 42 degrees heat shock for 60s, ice bath for 2min, add 900μl of antibiotic-free LB medium, and culture at 180rpm shaking for 45min; 4. Centrifuge at 6000rpm for 3min, leave 100μl of supernatant to resuspend, and then coat it on the corresponding resistant LB plate; 5.After inverting overnight, if there are too many colonies, the plasmid will be diluted for transformation, and if there are no colonies, 10ul plasmid will be taken for transformation; 6. Pick single colonies into the corresponding resistant LB liquid medium overnight shaking culture, and extract the plasmid as needed. Ⅱ. Liquid plasmid (minus 20 degrees storage, shelf life 90 days, ice pack transportation). 1. After receiving the liquid plasmid, please shake the preservation tube so that the liquid plasmid does not adsorb on the tube wall; 2.Take an appropriate amount of plasmid according to the experimental needs, and store the rest in a minus 20 degree freezer. Ⅲ. Glycerol strains (stored at minus 80 degrees, shelf life of 90 days, ice pack transportation). 1. After receiving the glycerol strain, please shake the preservation tube so that the bacterial solution does not adsorb on the tube wall; 2. Use the inoculation loop to mark the line in four areas on the solid plate, and add the corresponding resistance if there is resistance; 3. Inverted culture in constant temperature incubator for 12~16h; 4. Be sure to pick a single colony and inoculate it in the liquid medium, and add the corresponding resistance if there is resistance; 5. Shake the shaker and culture overnight, and culture the strains according to the needs of the experiment.
请按 Ctrl + D 键将本页加入收藏夹。
Mac 用户请按 ⌘ + D 键。
