Abstract

The vast majority of clinical samples are still archived using a decades-old technique of fixation with formaldehyde and embedding in paraffin wax to generate so-called formalin-fixed paraffin-embedded (FFPE) tissue blocks. This process of fixation and embedding, as well as long-term storage, can result in severe degradation of nucleic acids in the tissue. The fixation process itself causes cross-linkage between nucleic acids and proteins, as well as covalent modification of nucleic acids by monomethylol (-CH 2 OH) addition to the bases. Millions of FFPE blocks form the foundation of many tissue archives worldwide, and are essential to clinical research for exploring disease pathways and developing novel therapies. The inherent challenges of obtaining useable nucleic acids from difficult sample types such as these can be overcome by using the latest technologies and procedures to provide meaningful results throughout the next-generation sequencing workflow, from initial extraction of DNA/RNA to sample quantification, sample quality control, and library preparation.

Full Text
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