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Nucleic Acids Research, Vol 24, Issue 17 3472-3473, Copyright © 1996 by Oxford University Press

ARTICLES

A fluorescence-labeling method for sequencing small RNA on polyacrylamide gel

TP Wu, KC Ruan and WY Liu
Shanghai Institute of Biochemistry, Academia Sinica, China.

A practical fluorescence-labeling method for sequencing small RNAs by the traditional 'direct read out' on polyacrylamide gel electrophoresis was established. The 3' terminus of RNA was oxidized into dialdehyde by sodium periodate and then labeled with fluorescein-5-thiosemicarbazide through the condensation reaction between carbazide and aldehyde. The fluorescence-labeled RNA was partially degraded enzymatically and fractionated by polyacrylamide gel electrophoresis. The fluorescent bands were visualised by ultraviolet photography. A partial sequence of yeast 5S rRNA was determined. The result indicates that this method can be used in sequencing small RNAs rapidly, conveniently and safely.
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