title: Fractionation of non-polyadenylated and ribosomal-free RNAs from mammalian cells.
abstract: Most of mRNAs and well-characterized long noncoding RNAs are shaped with 5' cap and 3' poly(A) tail. Thereby, conventional transcriptome analysis typically involved the enrichment of poly(A)+ RNAs by oligo(dT) selection. However, accumulated lines of evidence suggest that there are many RNA transcripts processed in alternative ways, which largely failed to be detected by oligo(dT) purification. Here, we describe an enrichment strategy to purify non-polyadenylated (poly(A)-/ribo-) RNAs from total RNAs by removal of poly(A)+ RNA transcripts and ribosomal RNAs. In the combination with high-throughput sequencing methods, this strategy has been successfully applied to identify the rich repertoire of non-polyadenylated RNAs in vivo.
authors.affiliation: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
authors.forename: Qing-Fei
authors.initials: QF
authors.lastname: Yin
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Chen
authors.forename: Li
authors.initials: L
authors.lastname: Yang
mesh_terms: D000818:Animals; D005591:Chemical Fractionation; D008322:Mammals; D008967:Molecular Biology / Q000379:methods*; D026723:Polyadenylation; D011786:Quality Control; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification*; D012335:RNA, Ribosomal / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D012335:RNA, Ribosomal; D012313:RNA