Can u suggest the bacterial culture name that gives me more amount of plasmid from 1.5ml culture ?And let me know if i should take any precautions while performing experiment to avoid contamination other than plasmid in the desired sample ?
LB media with a supplementation of 0.1-0.2% Glucose is the best media. If plasmid are COLE1 in origin then use chloroamphencol to amplify the plasmid DNA and make sure to add chloroamphencol only after the culture has reached a good OD of 1.0
You need one of the common strains like XL1-Blue or DH5alpha (see: http://openwetware.org/wiki/E._coli_genotypes). They allow you to amplify large amounts of plasmid and blue/white screening. From a high copy number plasmid like the above mentioned pUC18 or with pBluescript II you will get around 10 µg of DNA fom a small culture. We just grow it in 2xYT with the respective antibiotica. Contamination is not a big issue. Just be carefully if you prepare plasmid of your gene of interest and try to do qPCR to quantify the same target from endogenous sources.
- Badges
- Science topic