Improved Screening of cDNAs Generated by mRNA Differential Display Enables the Selection of True Positives and the Isolation of Weakly Expressed Messages - Archive ouverte HAL Access content directly
Journal Articles Plant Molecular Biology Reporter Year : 1997

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

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 low abundance differential clones. Following cloning, a second reverse Northern combined with partial DNA sequencing and RT-PCR detection allows isolation of all differential cDNAs including very low abundance 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.
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hal-03607922 , version 1 (14-03-2022)

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Hicham Zegzouti, Christel Marty, Brian Jones, Thomas Bouquin, Alain Latché, et al.. Improved Screening of cDNAs Generated by mRNA Differential Display Enables the Selection of True Positives and the Isolation of Weakly Expressed Messages. Plant Molecular Biology Reporter, 1997, 1 (3), pp.236-245. ⟨10.1023/A:1007482318668⟩. ⟨hal-03607922⟩
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