Methods for the purification and detection of single nucleotide KRAS mutations on extrachromosomal circular DNA in human plasma.

ABSTRACT:

Backgrounds

Despite recent advances, many cancers are still detected too late for curative treatment. There is, therefore, a need for the development of new diagnostic methods and biomarkers. One approach may arise from the detection of extrachromosomal circular DNA (eccDNA), which is part of cell-free DNA in human plasma.

Aims

First, we assessed and compared two methods for the purification of eccDNA from plasma. Second, we tested for an easy diagnostic application of eccDNA liquid biopsy-based assays.

Materials & methods

For the comparison we tested a solid-phase silica purification method and a phenol/chloroform method with salt precipitation. For the diagnostic application of eccDNA we developed and tested a qPCR primer-based SNP detection system, for the detectio

SUBMITTER: Bollehuus Hansen L 

PROVIDER: S-EPMC10523981 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature