Northern blot analysis of simple repetitive sequence transcription in plants

Electrophoresis. 1996 Jul;17(7):1183-9. doi: 10.1002/elps.1150170702.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Northern*
  • Nucleic Acid Hybridization
  • Oligonucleotide Probes
  • Plants / genetics*
  • Poly A / metabolism
  • RNA, Messenger / analysis
  • RNA, Plant / analysis*
  • Repetitive Sequences, Nucleic Acid*

Substances

  • Oligonucleotide Probes
  • RNA, Messenger
  • RNA, Plant
  • Poly A