title: Identification of microRNAs and other small regulatory RNAs using cDNA library sequencing.
abstract: Distinct classes of small RNAs, 20-32 nucleotides long, play important regulatory roles for diverse cellular processes. It is therefore important to identify and quantify small RNAs as a function of development, tissue and cell type, in normal and disease states. Here we describe methods to prepare cDNA libraries from pools of small RNAs isolated from organisms, tissues or cells. These methods enable the identification of new members or new classes of small RNAs, and they are also suitable to obtain miRNA expression profiles based on clone count frequencies. This protocol includes the use of new deep sequencing methods (454/Roche and Solexa) to facilitate the characterization of diverse sequence pools of small RNAs.
authors.affiliation: Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, 1230 York Avenue, Box 186, New York, NY 10065, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Hafner
authors.forename: Pablo
authors.initials: P
authors.lastname: Landgraf
authors.forename: Janos
authors.initials: J
authors.lastname: Ludwig
authors.forename: Amanda
authors.initials: A
authors.lastname: Rice
authors.forename: Tolulope
authors.initials: T
authors.lastname: Ojo
authors.forename: Carolina
authors.initials: C
authors.lastname: Lin
authors.forename: Daniel
authors.initials: D
authors.lastname: Holoch
authors.forename: Cindy
authors.initials: C
authors.lastname: Lim
authors.forename: Thomas
authors.initials: T
authors.lastname: Tuschl
mesh_terms: D000818:Animals; D015723:Gene Library; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000302:isolation & purification; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D038621:Regulatory Sequences, Ribonucleic Acid / Q000235:genetics*
chemical_list: D035683:MicroRNAs; D038621:Regulatory Sequences, Ribonucleic Acid