NZYGigaprep

NZYGigaprep kit is designed for the rapid, large-scale preparation of highly pure plasmid DNA. The kit was optimized to obtain typically 2-10 milligrams of DNA from high-copy number plasmids or 500 micrograms to 2 milligrams of the nucleic acid from low-copy number plasmids, all originating from recombinant from recombinant Escherichia coli strains. Our kit streamlines the process with a modified alkaline/SDS lysis procedure. This procedure primes the bacterial cell pellet for plasmid purification. It involves denaturing both chromosomal and plasmid DNA under alkaline conditions. Next, the introduction of potassium acetate initiates the formation of a precipitate that contains chromosomal DNA and other cellular components. Simultaneously, the potassium acetate buffer neutralizes the lysate, allowing plasmid DNA to return to its native supercoiled state, keeping it in solution. After equilibrating the column with equilibration buffer, plasmid DNA selectively binds to the anion-exchange resin. With a thorough column wash, the plasmid DNA is efficiently purified and ready for elution. A simple precipitation step allows you to collect the eluted DNA, which can then be readily dissolved in TE buffer for subsequent use. The purified DNA is well-suited for the most demanding molecular biology applications, including transfection, in vitro transcription, automated fluorescent or manual sequencing, cloning, PCR, and hybridization. To isolate DNA from low-copy number plasmids, BACs or cosmids, double the volumes of Buffers M1, M2 and M3 (provided as NZY Buffers M for Midi, Maxi and Gigaprep kits, cat. nº. MB47701).