The EXTRACTME PLASMID DNA kit is designed for the rapid and efficient purification of high quality plasmid DNA from recombinant Escherichia coli strains. The isolation protocol and buffer formulations were optimised for high isolation efficiency and DNA purity. The product is intended for research use only.
The DNA purification procedure utilises spin minicolumns with membranes which efficiently and selectively bind nucleic acids. In the first isolation step, the plasmid DNA is released from bacterial cells by alkaline lysis. Then the lysate is neutralized and all the cell residues along with the proteins and genomic DNA are removed in the centrifugation step. The lysate is applied to the purification column membrane and the DNA is bound. The two-step washing stage effectively removes impurities and enzyme inhibitors. The purified plasmid DNA is eluted using a low ionic strength buffer (Elution Buffer) or water (pH 7.0-9.0) and may be used 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.
The quality of each production batch (LOT) of the EXTRACTME PLASMID DNA kit is tested using standard QC procedures. The purified DNA concentration, quality and stability are evaluated by gel electrophoresis and spectrophotometer. In addition, the functional quality is tested by qPCR and digestion with restriction enzymes.