HiPurA Endotoxin free Plasmid DNA Miniprep Purification Kit is supplied by HiMedia. HiPurA Endotoxin free Plasmid DNA Miniprep Purification Kit provides a fast and easy method for purification of endotoxin free plasmid DNA from E.coli host strains. The DNA purification procedure using the miniprep spin columns comprises of three steps viz. adsorption of DNA to the membrane, removal of residual contaminants and elution of pure plasmid DNA. HiMedias HiElute Miniprep Spin Column (Capped) 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 Concentrate (HBC)

Wash Solution (HPB)

Wash Solution Concentrate (HPE)

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

RNase A Solution (20 mg/ml)

Micro Centrifuge Tube-B (1.5 ml)

HiElute Miniprep Spin Column (Capped) [in DBCA016 Collection Tube]

Collection Tube (Uncapped), Polypropylene (2.0 ml)

Collection Tube, Polypropylene (2.0 ml)

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 Miniprep 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 onto 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 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 20 g of high- or low- copy plasmid DNA from 1-5 ml of E. coli culture per isolation procedure.