title: Analysis of Piwi-loaded small RNAs in Tetrahymena.
abstract: Scan RNAs (scnRNAs) are developmentally regulated siRNAs of ~26-32 nucleotides in length that are involved in programmed DNA elimination in Tetrahymena. scnRNAs are loaded onto the Piwi-related protein Twi1p and 2'-O-methylated at their 3' termini. We describe two alternative strategies for analyzing the Twi1p-loaded scnRNAs: preparation of loaded scnRNAs by immuno-purification of the Twi1p-scnRNA complex and exclusion of non-methylated scnRNAs during cDNA library construction using periodate oxidation.
authors.affiliation: Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Noto
authors.forename: Henriette M
authors.initials: HM
authors.lastname: Kurth
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Mochizuki
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015723:Gene Library; D047468:Immunoprecipitation; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000378:metabolism; D016133:Polymerase Chain Reaction; D016053:RNA, Protozoan / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D034741:RNA, Small Interfering / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D048348:Reverse Transcription; D013768:Tetrahymena / Q000235:genetics*
chemical_list: D060565:Argonaute Proteins; D018076:DNA, Complementary; D016053:RNA, Protozoan; D034741:RNA, Small Interfering; D010504:Periodic Acid; C009288:metaperiodate