How do you manufacture your Q-Seq HDx Reference Standards?

Our Q-Seq DNA HDx Reference Standards are manufactured by extracting genomic DNA from well-characterized, clonal human cell lines. The genomic DNA from multiple cell lines is stoichiometrically mixed to generate the desired multiplex of mutations. These variants are quantitatively verified using droplet-digital PCR at the loci of interest. For Q-Seq FFPE HDx Reference Standards, cell lines are blended to generate the desired mutation multiplex, and these are mildly-formalin fixed and then homogenously embedded in paraffin wax. The FFPE block is sectioned to provide individual FFPE curls, which may be extracted to yield ��400 ng of DNA*. *Yield depends on FFPE extraction method; our internal extraction methods utilize Promega's Maxwell LEV Plus FFPE extraction kit. Importantly, the cell lines used for manufacturing Q-Seq HDx Reference Standards also carry endogenous mutations, and these mutations may be exploited to develop a larger panel covering a number of mutations in one sample.