title: Barcoded cDNA library preparation for small RNA profiling by next-generation sequencing.
abstract: The characterization of post-transcriptional gene regulation by small regulatory (20-30 nt) RNAs, particularly miRNAs and piRNAs, has become a major focus of research in recent years. A prerequisite for characterizing small RNAs is their identification and quantification across different developmental stages, and in normal and disease tissues, as well as model cell lines. Here we present a step-by-step protocol for generating barcoded small RNA cDNA libraries compatible with Illumina HiSeq sequencing, thereby facilitating miRNA and other small RNA profiling of large sample collections.
authors.affiliation: Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, Box 186, NY 10065, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Hafner
authors.forename: Neil
authors.initials: N
authors.lastname: Renwick
authors.forename: Thalia A
authors.initials: TA
authors.lastname: Farazi
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Mihailović
authors.forename: John T G
authors.initials: JT
authors.lastname: Pena
authors.forename: Thomas
authors.initials: T
authors.lastname: Tuschl
mesh_terms: D001483:Base Sequence; D020869:Gene Expression Profiling / Q000379:methods; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics; D017423:Sequence Analysis, RNA / Q000379:methods; D013048:Specimen Handling
chemical_list: D035683:MicroRNAs; D034741:RNA, Small Interfering