The DNA purification procedure utilizes spin 96-minicolumns plates 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 separated with the use of Plasmid Filter Plate. Then the lysate is applied to the purification minicolumn 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.
Bacterial broth culture
Approx. 20 μg DNA per well
Approx. 40 minutes for purification using centrifuge or vacuum manifold
A260/A280 ratio =1.7 – 1.9