CIMac™ pDNA-0.3 Analytical Columns (Pores 1.4 μm) were introduced by BIA Separations d.o.o. in 2010. In the following years they were recognised by the biotechnological and analytical community as excellent choice for fast and exact plasmid DNA (pDNA) monitoring and quantification. Due to high sensitivity of the column to small changes in chromatographic conditions we publish this technical note, how to exploit the column’s excellent chromatographic characteristics to the highest possible extent.
A standard testing protocol for the columns is analytical separations of open circular (oc) and supercoiled (sc) pDNA isoforms of a 4.7 kbp large plasmid DNA molecule. The concentration of the pDNA sample used was 23μg/mL. The sample contained between 21 and 22% of the oc pDNA isoform and an exact amount of uracil as unbound tracer. The separation of the isoforms was performed in a linear gradient of NaCl with the temperature of the whole system controlled to 15.0±0.5 °C. The typical experimental conditions together with an example of chromatogram are shown in Figure 1.