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Northern blot analysis of simple repetitive sequence transcription in plants
Author(s) -
Gortner Götz,
Pfenninger Markus,
Kahl Günter,
Weising Kurt
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150170702
Subject(s) - northern blot , southern blot , biology , oligonucleotide , microbiology and biotechnology , repeated sequence , oligomer restriction , blot , nucleic acid thermodynamics , in situ hybridization , rna , dot blot , western blot , sequence analysis , messenger rna , gene , genetics , genome
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.