cDNA Library Generation for the Analysis of Small RNAs by High-Throughput Sequencing

• DOI: 10.1007/978-1-4939-2547-6_13
• Publication Date: 2015-03-19T09:58:16Z
• Authors: Jennifer Gebetsbe...; Roger Fricker; Norbert Polacek
• Keywords: 0301 basic medicine; 0303 health sciences; 03 medical and health sciences; 13. Climate action; Trypanosoma brucei brucei; High-Throughput Nucleotide Sequencing; RNA, Small Untranslated; Gene Library

The RNome of a cell is highly diverse and consists besides messenger RNAs (mRNAs), transfer RNAs (tRNAs), and ribosomal RNAs (rRNAs) also of other small and long transcript entities without apparent coding potential. This class of molecules, commonly referred to as non-protein-coding RNAs (ncRNAs), is involved in regulating numerous biological processes and thought to contribute to cellular complexity. Therefore, much effort is put into their identification and further functional characterization. Here we provide a cost-effective and reliable method for cDNA library construction of small RNAs in the size range of 20-500 residues. The effectiveness of the described method is demonstrated by the analysis of ribosome-associated small RNAs in the eukaryotic model organism Trypanosoma brucei.