Amplicon Sequencing from FFPE Tissues on the MiSeq® System

This application note presents a simple and fast workflow for accurate detection of rare variants even from degraded DNA. It describes sequencing indexed amplicons of KRAS oncogene exon 2 region from genomic DNA extracted from formalin-fixed, paraffin embedded (FFPE) paired tumor/normal samples. From a single MiSeq run, coverages in excess of 150,000-fold were observed for each amplicon, sufficient to detect variants close to the limit of detection (~0.5%), even from highly degraded DNA.