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Journal Article

RNA sequencing using fluorescent-labeled dideoxynucleotides and automated fluorescence detection.

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Bauer,  G. J.
Abteilung Biochemische Kinetik, MPI for biophysical chemistry, Max Planck Society;

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603041.pdf
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Citation

Bauer, G. J. (1990). RNA sequencing using fluorescent-labeled dideoxynucleotides and automated fluorescence detection. Nucleic Acids Research, 18(4), 879-884.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0013-0E38-3
Abstract
Although dideoxy terminated sequencing of RNA, using reverse transcriptase and oligodeoxynucleotide primers, is now a well established method, the accuracy is limited by sequence ambiguities due to unspecific chain termination events. A protocol is described which circumvents these ambiguities by using fluorescence labels tagged to dideoxynucleotides. Only chain terminations caused by dideoxynucleotides were detected while premature terminated cDNA's remain undetectable. In addition, the remaining multiple signals at nucleotide positions can be assigned to sequence heterogeneities within the RNA sequence to be determined.