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Improved Screening of cDNAs Generated by mRNA Differential Display Enables the Selection of True Positives and the Isolation of Weakly Expressed Messages

Zegzouti, Hicham and Marty, Christel and Jones, Brian and Bouquin, Thomas and Latché, Alain and Pech, Jean-Claude and Bouzayen, Mondher Improved Screening of cDNAs Generated by mRNA Differential Display Enables the Selection of True Positives and the Isolation of Weakly Expressed Messages. (1997) Plant Molecular Biology Reporter, vol. 1 (n° 3). pp. 236-245. ISSN 0735-9640

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Official URL: http://dx.doi.org/10.1023/A:1007482318668

Abstract

The high percentage of false positives generated by differential display (as high as 85%) has previously limited the potential of the method. This report describes an efficient methodology that enables false positives to be discarded prior to cloning, via reverse Northern analysis. This first step of the screening also allows the detection of putative lowabundance differential clones. Following cloning, a second reverseNorthern combined with partial DNA sequencing and RT-PCR detection allows isolation of all differential cDNAs including very lowabundance clones. Use of the sequential screening procedure described here led to the isolation of novel tomato genes responding to the plant hormone ethylene while minimising labor and materials input.

Item Type:Article
Additional Information:Thanks to Kluwer editor. The definitive version is available at http://www.springerlink.com The original PDF of the article can be found at Plant Molecular Biology Reporter website : http://www.springerlink.com/content/112114/
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Institut National de la Recherche Agronomique - INRA
Université de Toulouse > Institut National Polytechnique de Toulouse - INPT
Laboratory name:
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Deposited By:Dabernat Séverine

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