Novel Technology for Enrichment of Cell-free Biomolecules and Size Fractionation of ccfDNA
Circulating DNA plays a significant role in prenatal screening. In the near future tumor derived ccfDNA will play an important role in cancer diagnostics and in cancer patient monitoring. How information delivered from ccfDNA and ccfRNA and applied to diagnostics tests are currently being explored. Nevertheless, the crucial step for efficient genetic analysis is the isolation of sufficient amounts of ccfNA even if the nucleic acid content is very low and it is highly fragmented.
Analytik Jena’s PME technology is a novel and simple technology for efficient enrichment of biomolecules e.g. circulating cell-free DNA (ccfDNA) up to 10 ml of sample. The technology captures the biomolecules with a polymer as the first step, followed by an efficient nucleic acids extraction
The PME free-circulating DNA Extraction Kit is based on a new, patent-pending technology called PME – Polymer Mediated Enrichment. The first step captures cell-free DNA in the entire sample by encapsulating it with the PME polymer. Afterwards this complex is collected as a pellet by centrifugation. The captured nucleic acid is dissolved in a special buffer thus significantly reducing the sample volume in the following extraction steps. Even when the amount of freely circulating DNA is very low and only small nucleic acid fragments (<100 nt) are available, the technology delivers fast reliable results. Subsequent downstream application such as digital PCR can be performed with the eluvate without any additional processing.
Additionally the PME technology offers the possibility for selective DNA fractionation. Based on this new method the total ccfDNA can be separated in a long chain DNA fraction and in a short chain DNA fraction. The new method allows to enrich the DNA fraction of interest as well as to remove undesirable DNA. The DNA fractionation method has been already applied for removal of maternal long chain DNA for prenatal screening and for removal of wild type long chain DNA for biomarker analysis.
The PME technology is a simple and efficient method for extraction ccfDNA from a range of biological samples.