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Jacqueline Chan, Juliette Forster, Aysel Heckel, Venu Pullabhatla, Dave Cook, Graham Speight



Formalin-fixed, paraffin-embedded (FFPE) storage is a standard method for archiving samples from solid tumours. It ensures the preservation of the ultrastructure of tissues and prevents degradation through formation of chemical links between macromolecules, for example between and within DNA molecules. FFPE samples contain a wealth of information which can be used to study cancer development and progression. Next generation sequencing (NGS) offers the capability of unlocking this information through the simultaneous study of multiple types of mutations in cancer-associated genes for a number of applications1. However, formalin treatment can significantly compromise the quality and amount of nucleic acids available for genomics research. As such it is technically challenging to examine the true genetic complexity present in a sample.

In this study DNA reference standards with different levels of formalin-induced damage were hybridised and sequenced with a SureSeq™ custom NGS panel in conjunction with the SureSeq FFPE DNA Repair Mix*. We assessed the impact of the repair mix on three levels of formalin compromised DNA (fcDNA) - ‘mild’, ‘moderate’ and ‘severe’, at 4 different DNA input amounts down to 10 ng. We then compared the uniformity and coverage of the enriched targets. We also assessed the concordance to the allele frequencies of the variants in the reference standards.

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