title: Small RNA Library Construction for Exosomal RNA from Biological Samples for the Ion Torrent PGM™ and Ion S5™ System.
abstract: Next-generation deep sequencing (NGS) technology represents a powerful and innovative approach to profile small RNA. Currently, there are a number of large-scale and benchtop sequencing platforms available on the market. Although each platform is relatively straightforward to operate, constructing cDNA libraries can be the most difficult part of the NGS workflow. Constructing quality libraries is essential to obtaining a successful sequencing run of high-quality reads and coverage. The quality and yield of RNA affect hybridization and ligation of sequencing adapters. In the field of biomarker discovery, there has been an interest in profiling exosomal RNA from biological fluids. However, very little RNA yield is obtained when extracting RNA from exosomes, thus making library construction difficult. Here, this protocol describes an optimized protocol for constructing small RNA libraries from low yields of RNA, in particular, extracted from exosomes isolated from biological fluids.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Melbourne, Melbourne, VIC, Australia.|Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science (LIMS), La Trobe University, Room 415, LIMS1, Bundoora, Melbourne, VIC, 3086, Australia.
authors.forename: Lesley
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, VIC, Australia. Andrew.hill@latrobe.edu.au.
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Hill
mesh_terms: D055354:Exosomes; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D035683:MicroRNAs / Q000235:genetics / Q000302:isolation & purification; D058727:RNA, Small Untranslated / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D035683:MicroRNAs; D058727:RNA, Small Untranslated
keywords: Biological fluids; Exosomes; Ion Torrent; Small RNA; Small RNA deep sequencing; miRNA