Principle of RNA sequencing

Klinische Monatsblätter für Augenheilkunde (2020)

RNA Sequencing of Formalin-Fixed and Paraffin-Embedded Tissue as a Complementary Method in Ophthalmopathology

Günther Schlunck, Stefaniya Boneva, Julian Wolf, Anja Schlecht, Thomas Reinhard, Claudia Auw-Hädrich, Clemens Lange



The high-throughput method of "Next Generation Sequencing" (NGS) allows cost-effective decoding of the nucleotide sequences of millions of RNA molecules in a sample. This makes it possible to determine the number of distinct RNA molecules in tissues or cells and to use these data to draw conclusions. The entirety of RNAs, in particular mRNAs (messenger RNAs) as potential precursors of proteins, provides a comprehensive insight into the functional state of the cells and tissues under investigation. In addition to cell cultures or unfixed tissue, formalin-fixed and paraffin-embedded (FFPE) tissue can also be analysed for this purpose using specific methods. In this overview, the methodological strategy and its application to the field of ophthalmic histopathology are presented.