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Nucleic Acids Res. 1997 May 15;25(10):2030-1.

Precise branch point mapping and quantification of splicing intermediates.

Author information

1
Department of Biology, Humboldt University, Chausseestrasse 117, 10115 Berlin, Germany. joerg=vogel@rz.hu-berlin.de

Abstract

Lariat intermediates of a group II intron were investigated via RT-PCR. Several reverse transcriptases appeared capable of reading through a branched nucleotide. A new method has been established that yields precise information about the location of the branch point within an intron. As an extension of our approach, antisense transcripts of the previously cloned PCR products were successfully used in RNase Protection Assays, providing a tool for quantification of splicing intermediates. Application of the method presented to other self-splicing introns as well as introns in nuclear pre-mRNAs is envisaged.

PMID:
9115373
PMCID:
PMC146694
DOI:
10.1093/nar/25.10.2030
[Indexed for MEDLINE]
Free PMC Article

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