title: Generation of a large catalog of unique transcripts for whole-genome expression analysis in nonmodel species.
abstract: Next-generation sequencing technologies have allowed new tools to be developed for transcriptome characterization. We describe here the methods for the preparation of a library from pooled RNAs for 454 sequencing of 3'-cDNA fragments. We also describe how the read sequences obtained can be used to generate, through de novo reads assembly, a large catalog of unique transcripts in organisms for which a comprehensive collection of transcripts or the complete genome sequence is not available. Finally, this catalog can be used efficiently to design oligonucleotide probes for setting up a comprehensive microarray for global analysis of gene expression.
authors.affiliation: Department of Biotechnology, University of Verona, Verona, Italy.
authors.forename: Diana
authors.initials: D
authors.lastname: Bellin
authors.forename: Alberto
authors.initials: A
authors.lastname: Ferrarini
authors.forename: Massimo
authors.initials: M
authors.lastname: Delledonne
mesh_terms: D018076:DNA, Complementary / Q000235:genetics; D030541:Databases, Genetic; D020869:Gene Expression Profiling; D015723:Gene Library; D016678:Genome; D046228:Microarray Analysis; D015345:Oligonucleotide Probes / Q000235:genetics; D012313:RNA / Q000032:analysis / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D027843:Vitis / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D015345:Oligonucleotide Probes; D018749:RNA, Plant; D012313:RNA