BAC DNA purification—NucleoBond Xtra BAC

NucleoBond Xtra BAC provides second-generation anion exchange for large-construct plasmid DNA. The high selectivity and specific characteristics of the optimized silica matrix make this anion exchange kit ideal for efficiently purifying very large constructs, such as P1, BACs, and PACs. For example, using the NucleoBond Xtra BAC kit, it takes just 75 minutes to isolate BAC DNA free of all types of RNA, proteins, metabolites, dyes, and carbohydrates. This kit completely removes impurities which can act as strong inhibitors for enzymatic processing, transfections, transcriptions, etc., even when present in small amounts.

LyseControl, a blue pH indicator reagent present in the lysis buffer used in the NucleoBond Xtra BAC procedure, ensures complete and efficient alkaline lysis, providing optimal yields. Upon addition of lysis buffer to the resuspended cells, the cell suspension turns blue. To avoid contamination with genomic DNA, the blue solution is gently inverted up to five times until a homogeneous and 100% colorless solution is achieved. After a five-minute incubation, neutralization buffer is added and LyseControl turns colorless. Traces of blue LyseControl indicate insufficient mixing. Using LyseControl ensures sufficient mixing and complete neutralization, which are necessary to renature plasmid DNA, precipitate proteins/gDNA, and remove SDS, which inhibits plasmid binding.