title: Northern blot analysis of simple repetitive sequence transcription in plants.
abstract: The presence of simple repetitive sequence motifs in RNA from various plant species was probed by Northern blot analysis. Hybridization of total, poly(A)(+)- and poly(A)(-)-RNA with microsatellite-complementary oligonucleotide probes revealed distinct bands with most but not all probe/species combinations, demonstrating the presence of di-, tri- and tetranucleotide repeat motifs in plant transcripts. Only trinucleotide repeat-derived hybridization signals were found to be enriched in the poly(A)(+)-fraction. The quality of Northern blot signals proved to be highly dependent on hybridization stringency. Thus, under the stringency conditions usually applied for oligonucleotide hybridization, some probes [(GT)8, (CAC)5, (TCC)5, and (CCTA)4] cross-hybridized to bands corresponding in size to 18S and/or 26S rRNA. Cross-hybridization to rRNA was significantly reduced at higher stringencies. These results stress the importance of carefully adjusting the hybridization conditions in Northern blot analysis of simple sequence transcripts.
authors.affiliation: Department of Biology, University of Frankfurt am Main, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Gortner
authors.forename: M
authors.initials: M
authors.lastname: Pfenninger
authors.forename: G
authors.initials: G
authors.lastname: Kahl
authors.forename: K
authors.initials: K
authors.lastname: Weising
mesh_terms: D015152:Blotting, Northern; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010944:Plants / Q000235:genetics*; D011061:Poly A / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis*; D012091:Repetitive Sequences, Nucleic Acid
chemical_list: D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D018749:RNA, Plant; D011061:Poly A