title: Identification of novel long noncoding RNA transcripts in male germ cells.
abstract: Emerging evidence from these studies suggested that the male germ cell transcriptome is more complex than previously envisioned. In addition to protein-coding genes, the transcriptome also encodes a significant number of nonprotein-coding transcripts. These noncoding (nc) RNAs appear to be involved in a variety of cellular activities, ranging from simple housekeeping to complex regulatory functions. A class of ncRNAs known as long ncRNAs (lncRNAs) were recently shown to be expressed in a developmentally regulated manner during brain and embryonic stem cell development. This protocol aims to predict and identify potential lncRNA candidates using Serial Analysis of Gene Expression (SAGE) data. We also illustrate how to validate the potential lncRNAs by expression analyses using real-time PCR and Northern Blot. Potential lncRNA candidates in male germ cells are identified using our previously established male germ cell SAGE database (GermSAGE).
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong, China. leetl@mail.nih.gov
authors.forename: Tin-Lap
authors.initials: TL
authors.lastname: Lee
authors.forename: Amy
authors.initials: A
authors.lastname: Xiao
authors.forename: Owen M
authors.initials: OM
authors.lastname: Rennert
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D019295:Computational Biology / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D008297:Male; D051379:Mice; D016366:Open Reading Frames; D022661:RNA, Untranslated / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D013093:Spermatogonia / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D022661:RNA, Untranslated