HiPurA Endotoxin free Plasmid DNA Maxiprep Purification Kit is supplied by HiMedia. HiPurA Endotoxin free Plasmid DNA Maxiprep Purification Kit provides a fast and easy method for purification of endotoxin free plasmid DNA. The DNA purification procedure using the maxiprep spin columns comprises of three steps viz. adsorption of DNA to the membrane, removal of residual contaminants and elution of pure plasmid DNA. HiMedias HiPure Maxiprep column format allows rapid processing of multiple samples. The columns have a high binding capacity and high-quality DNA is obtained. The endotoxin free plasmid DNA obtained is compatible with downstream applications such as transfection, automated sequencing etc. This kit contains:

Resuspension Solution (HP1)

Lysis Solution (HP2)

Neutral Solution M (NSM)

Endotoxin Removal Solution (ERS)

Binding Solution (HB)

Wash Solution M (WSM)

Wash Solution Concentrate (HPE)

Elution Buffer (ET) [10 mM Tris-Cl, pH 8.5]

RNase A Solution (20 mg/ml)

HiPure Midiprep Spin Column (in PW143 Collection Tube)

HiPure Midiprep Syringe Filter

Collection Tube (50 ml conical)

Endotoxins, also known as lipopolysaccharides, are present on the cell membrane of Gram negative bacteria such as E.coli. During the lysis step of plasmid purification, endotoxins are released, which significantly reduce transfection efficiencies in endotoxin sensitive cell lines. Endotoxins represent a non-controllable variable in transfection experiment setup, influencing the outcome and reproducibility of results and making them difficult to compare and interpret. In gene therapy research, endotoxins can interfere by causing endotoxic shock syndrome and activation of the complement cascade. The HiPurA Endotoxin free Plasmid DNA Maxiprep Purification Kit integrates an efficient endotoxin removal step into plasmid purification procedure. Cells are lysed and endotoxins are selectively precipitated in a specialized buffer. The resultant lysate is then applied on to the silica column for binding of the DNA molecules in the presence of high salt concentration. The adsorbed DNA is washed to remove contaminants, and the pure endotoxin free plasmid DNA is eluted in Elution Buffer. The purified plasmid DNA can be used for immediate use in all molecular biology procedures such as transfection, automated sequencing and other downstream applications. The kit utilizes an advanced silica-based membrane technology in the form of a convenient spin column which helps to recover upto 500 g of high- or low- copy plasmid DNA from upto 500 ml of E. coli culture per isolation procedure. The yield depends on plasmid copy number, size of insert, host strain, culture medium and culture volumes. The endotoxin levels are efficiently reduced to less than 0.1 EU/g.