The NucleoBond BAC 100 Kit is designed for purifying large DNA fragments such as cosmids, bacteriophage P1 clones, P1-derived artificial chromosomes (PACs), and bacterial artificial chromosomes (BACs), without phenol/chloroform extraction. The procedure takes less than 1 hour to isolate BAC DNA free from any kind of RNA, proteins, metabolites, dyes, or carbohydrates. After purification, all nucleic acids show absorbance ratios of A260/A280 as high as 1.8–2.0.
Low amounts of impurities, which often act as strong inhibitors for enzymatic processing, sequencing, transfections, transcriptions, etc., are removed completely.
NucleoBond Folded Filters, which are specially designed to eliminate the centrifugation step after the alkaline lysis step, are provided with the kit. Using the filters reduces hands-on-time and eliminates plastic waste common to other filtration systems. In approximately 10 minutes of unattended operation, complete removal of SDS and cellular debris from plasmid samples is achieved. NucleoBond Folded Filters do not induce shearing of large DNA constructs, such as PACs or BACs.
The patented anion exchange resin in the NucleoBond column is known to provide high-purity DNA equivalent or superior to that obtained by two successive rounds of CsCl ultracentrifugation. The plasmid purification procedure is performed in the absence of toxic material such as phenol, chloroform, ethidium bromide, or CsCl.