High Purity Plasmid MiniPrep Kit

Supplier Info.

Biotekeâ€™s Plasmid MiniPrep Kit is designed for the rapid preparation of plasmid DNA from small batch cultures of Escherichia coli. The plasmid DNA is preferentially purified from other lar components such as genomic DNA and RNA. The Plasmid MiniPrep Kit is able to purify plasmids up to 13,000 bp in size, and the typical purification yield is up to 20 ïg from 1.5 mL of bacterial culture. The purified DNA is fully digestible with all restriction enzymes tested, and is completely compatible with manual or automated sequencing to achieve 95-100% accuracy.

Purification is based on spin column chromatogra. The process for the isolation of plasmid DNA involves first pelleting 1.5 mL of an overnight culture of E. coli harbouring the plasmid of interest using centrifugation. The pellet is then resuspended in the provided Resuspension Solution AZ, which contains RNase A. Lysis Buffer N is then added to the sample in order to assist in the lysis of the bacterial . Next, Buffer TN is added to the sample which will neutralize the sample and cause precipitation of the proteins and genomic DNA that is present. The resulting suspension is spun down, and the clarified lysate containing the plasmid DNA is then applied to a provided column through centrifugation.

Biotekeâ€™s column binds DNA in a manner that depends on ionic concentrations, thus the DNA will bind to the column while most of the RNA, proteins and other contaminants will either flowthrough or be retained on top. The bound DNA is then washed twice using the provided Wash Solution E in order to remove any remaining impurities, and the purified PCR product is eluted with the Elution Buffer K.

The Buffer TN contains guanidine hydrochloride, and should be handled with care. Guanidine hydrochloride forms highly reactive compounds when combined with bleach, thus care must be taken to properly dispose of any of this solution.