 This method has the limitation of not being able to detect larger structural variant present in ctDNA and for this reason massively parallel next generation sequencing is also used to determine ctDNA content in cfDNA

Massively parallel sequencing (MPS) has allowed the deep sequencing of cfDNA.This deep sequencing is required to detect mutant ctDNA present in low concentrations in the plasma.Two main sequencing techniques are typically used for analysis of mutant cfDNA; PCR amplicon sequencing and hybrid capture sequencing.