The High-Speed Plasmid Mini Kit (10-50 Kb) was designed for plasmid and cosmid DNA purification from 1-4 ml of cultured bacterial cells. This plasmid kit is optimized for isolating 10-50 kb plasmid DNA using a miniprep spin column system. High-Speed Plasmid Mini Kit (10-50 Kb) uses a modified alkaline lysis method and RNase treatment to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. In the presence of chaotropic salt, plasmid DNA in the lysate binds to the glass fiber matrix of the plasmid spin column. Contaminants are removed with a Wash Buffer (containing ethanol) and the purified plasmid DNA is eluted by a low salt Elution Buffer or TE. Typical yields are 20-30 μg for high-copy number plasmid or 3-10 μg for low-copy number plasmid from 4 ml of cultured bacterial cells. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 15 minutes. The purified plasmid DNA is ready for use in Restriction Enzyme Digestion, Ligation, PCR, and sequencing reactions.
Advantages (Cat. # PDL100, PDL300)
Purify 10-50 kb plasmid DNA within 15 minutes!
High Yield:up to 30 μg of plasmid DNA or cosmid DNA
Sample:1-4 ml of bacterial cells (10-50 kb plasmid)
Spin Columns: glass fiber membrane optimized for large plasmd DNA purification
Storage:room temperature (15-25ºC) for up to 1 year
The quality of the High-Speed Plasmid Mini Kit (10-50 Kb) is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. 5 µl aliquots of a 100 µl eluate of purified super coiled plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing a 17.4 kb low copy plasmid pBluescript (OD600 = 3.5 U/ml) are used in EcoRI digestion and analyzed by electrophoresis on a 0.8% agarose gel.
