QUANTIFICATION Circulating DNA

Quantification of cfDNA is ideally carried out by qPCR or capillary electrophoresis since common methods such as absorption measurement or fluorescent dye based quantification might lead to false results due to low DNA concentration.

The total cf DNA isolated can be quantified using the Cell-free human DNA detc-qPCR Test designed to  target a conserve sequence region of a gene repeated more than a hundred times in the human genome.

Are individuals ready-to-use tubes containing all the components needed to perform the quantitative PCR assay.

Specifications:

Real-time PCR amplification plot for cfhDNA dtec-qPCR Test (red) targeting a “non-truncated”      multi-copy gene and compared to a monocopy target (blue), using a human genomic DNA as a standard. Due to the presence of multiple copies of the selected target, sensibility is increased 2 logs (100 times) for the cfhDNA dtec-qPCR Test. Same increased signal is observed for the purified cell-free DNA samples employed for cell-free DNA quantification.

Quantification of cf-DNA from plasma

Blood samples were collected from 8 patients (samples 1 to 8)  with breast cancer and healthy controls. 2 samples were used for healthy individuals (sample 9 and 10) and 2 samples of healthy individuals were spiked with 150 ng (sample 11) and 300 ng (sample 12) of human genomic DNA.

Circulating cell-free DNA was extracted from 3 ml of plasma following DANAGENE Circulating DNA Kit protocol and quantified using the Cell-free human DNA detc-qPCR Test.

We successfully detected increased concentrations of circulating cell free-DNA in all cancer patients.