EXTRACTME PLASMID MAXI KIT, 25 preps supplied by BLIRT is intended to ensure efficient purification of high quality plasmid DNA from 200-1000 ml of cultured bacterial cells. The kits are suitable for anion-exchange resins that allow extraction of ultrapure, transfectiongrade pDNA that is utilized in demanding applications. The protocol of isolation and buffer formulations was optimized to ensure high efficiency of isolation and purity of pDNA. The process for plasmid DNA purification uses pre-packed anion-exchange resin columns which bind nucleic acids efficiently and selectively. The 1st isolation step involves the release of pDNA from bacterial cells by alkaline lysis. This is followed by the neutralization of lysate and separation of all cell residues along with proteins and genomic DNA by centrifugation. This alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. Lysate is applied to purification column membrane with the equilibrated resin and DNA is bound. The two-step washing stage adequately eliminates pollutants and enzyme inhibitors. The elution of purified DNA is done with the help of either a low ionic strength buffer (Elution Buffer) or water (pH 7.09.0). And it may be utilized directly in all downstream applications such as PCR, qPCR, Southern blotting, DNA sequencing, enzymatic restriction, ligation and so forth, or stored until ready to use.
Product name - EXTRACTME PLASMID MAXI KIT, 25 preps
Catalog No - EM18-025
Pack size: 25 preps
1 SAMPLE MATERIAL
200-600 ml (high-copy number plasmids)
400-1000 ml (medium- and low-copy number plasmids).
1.0-1.5 mg of transfection-grade pDNA from 400 ml of cultured bacterial cells
3. TIME REQUIRED
90 min for EM18 and 120 min for EM19
additional ~60 min for DNA precipitation
4. DNA PURITY
A260/A280 ratio = 1.7 1.9