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Hybridizationin situ of whole-mount messenger RNA in plants

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Abstract

A procedure is presented for the detection of mRNA in whole-mount preparations of youngArabidopsis seedlings using digoxigenin (DIG)-labeled RNA probes. It includes tissue preservation with formaldehyde, permeabilization with polyoxyethy lenesorbitan (Tween 20), DMSO, and heptane. Hybridization signal is detected using colloidal gold or alkaline-phosphatase-conjugated anti-DIG antibodies.

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Abbreviations

DIC:

differential interference contrast

DIG:

digoxigenin

IAA:

indol-3-acetic acid

ISH:

in-situ hybridization

SOD:

superoxide dismutase

Tween:

polyoxyethylenesorbitan

WISH:

whole-mountin situ hybridization

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Authors and Affiliations

  1. Laboratorium voor Genetica, Universiteit Gent, K.L. Ledeganckstraat 35, B-9000, Gent, Belgium

    Janice de Almeida Engler & Marc Van Montagu

  2. Laboratoire Associé de l'Institut National de la Recherche Agronomique, (France)

    Gilbert Engler

Authors
  1. Janice de Almeida Engler
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  2. Marc Van Montagu
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  3. Gilbert Engler
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Correspondence to Marc Van Montagu.

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de Almeida Engler, J., Van Montagu, M. & Engler, G. Hybridizationin situ of whole-mount messenger RNA in plants. Plant Mol Biol Rep 12, 321–331 (1994). https://doi.org/10.1007/BF02669275

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