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Construction and Analysis of Full-Length and Normalized cDNA Libraries from Citrus

  • M. Carmen Marques
  • Miguel A. Perez-Amador
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 815)

Abstract

We have developed an integrated method to generate a normalized cDNA collection enriched in full-length and rare transcripts from citrus, using different species and multiple tissues and developmental stages. Interpretation of ever-increasing raw sequence information generated by modern genome sequencing technologies faces multiple challenges, such as gene function analysis and genome annotation. In this regard, the availability of full-length cDNA clones facilitates functional analysis of the corresponding genes enabling manipulation of their expression and the generation of a variety of tagged versions of the native protein. The development of full-length cDNA sequences has the power to improve the quality of genome annotation, as well as provide tools for functional characterization of genes.

Key words

Library cDNA Citrus Full-length Normalized SMART DSN nuclease Gateway technology 

Notes

Acknowledgments

The authors would like to thank to all participants in the Spanish Citrus Functional Genomic Project, specially to Drs. Javier Forment, Jose Gadea, and Vicente Conejero. This work was funded by grants from the Spanish Government GEN2001-4885-CO5-01 and GEN2001-4885-CO5-02.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Instituto de Biología Molecular y Celular de Plantas (IBMCP), Universidad Politécnica de Valencia (UPV) and Consejo Superior de Investigaciones Científicas (CSIC)ValenciaSpain
  2. 2.Pérez-AmadorInstituto de Biología Molecular y Celular de Plantas, CSIC-Universidad Politécnica de Valencia, Ciudad Politécnica de la InnovaciónValenciaSpain

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