Clinical Chemistry
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Clinical Chemistry 44: 2249-2255, 1998;
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Right arrow Molecular Diagnostics and Genetics
(Clinical Chemistry. 1998;44:2249-2255.)
© 1998 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

RNA chip: quality assessment of RNA by microchannel linear gel electrophoresis in injection-molded plastic chips

Mieko Ogura1, Yoichi Agata2, Kenji Watanabe2, Randy M. McCormick3, Yohei Hamaguchi4, Yoshimasa Aso4, and Masato Mitsuhashi1,4,a

1 Hitachi Chemical Research Center, Irvine, CA 92612.

2 Goshomiya Works, Hitachi Chemical Company, Ltd., Shimodate 308, Japan.

3 Soane BioSciences, Hayward, CA 94545-3716.

4 Department of Pathology, University of California, Irvine, CA 92697.
a Address correspondence to this author at: 1003 Health Sciences Road West, Irvine, CA 92612. Fax 714-725-2727; e-mail mmitsuha{at}uci.edu.

Two major components of rRNA (18S and 28S rRNA) were separated by electrophoresis in injection-molded acrylic chips with a microchannel 100 µm in width, 40 µm in depth, and with 1 cm of separation distance. Microchannels were filled with 4 g/L hydroxypropylmethylcellulose as sieving polymer and 5 mg/L ethidium bromide for RNA staining. The fluorescent signals were detected by a fluorescent microscope equipped with a photometer and 590 nm emission filter. The assay is rapid (<3 min), reproducible, RNase-free, and requires only 1–2 µL of sample. The detection limit was ~10 mg/L (10 ng/µL), 100-fold lower than that for conventional agarose gel electrophoresis. Because only 0.1 nL of the loaded sample was used for electrophoresis, the detectable peaks of rRNA in the separation were derived from less RNA than in a single cell. Because the quality of RNA is critical for RNA-related diagnostic tests, disposable plastic chips will be useful for quality assessment of RNA.




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