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Enhanced detection of tRNA isoacceptors ... l oligonucleotide hybridization


Enhanced detection of tRNA isoacceptors by combinatorial oligonucleotide hybridization.

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A method that greatly enhances the detection of tRNA by oligodeoxyribonucleotide probe hybridization has been developed. Because highly structured tRNA regions often preclude heteroduplex formation, we have tested the ability of cold oligodeoxyribonucleotides called unfolders to disrupt the tRNA secondary/tertiary structures and promote hybridization of a second labeled oligonucleotide complementary to the anticodon loop. Here we show that an excess of unfolders in the pre/hybridization reaction can enhance a barely detectable hybridization signal by more than 200-fold without affecting probe specificity. This sensitive assay makes it possible to easily study and monitor changes in tRNA isoacceptor expression.


Buvoli A, Buvoli M, Leinwand LA

RNA (New York, N.Y.)

2000-06-01 00:00

6

6

912-8

Base Sequence,Cell Line,Combinatorial Chemistry Techniques,Humans,Molecular Sequence Data,Nucleic Acid Conformation,Nucleic Acid Hybridization,Oligodeoxyribonucleotides,Oligonucleotide Probes,RNA, Transfer, Ser,Oligodeoxyribonucleotides,Oligonucleotide Probes,RNA, Transfer, Ser

Department of Molecular, Cellular, and Developmental Biology, University of Colorado at Boulder, 80309, USA

RNA

NHLBI HL50560

1355-8382




710

True

10864048

Massimo Buvoli
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