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Development of an Efficient Inverse PCR Method for Isolating Gene Tags from T-DNA Insertional Mutants in Rice

  • Sung-Ryul Kim
  • Jong-Seong Jeon
  • Gynheung An
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 678)

Abstract

The central goal of current genomics research in plants, as in other organisms, is to elucidate the functions of every gene. Insertional mutagenesis using known DNA sequences such as T-DNA is a powerful tool in functional genomics. Development of efficient methods for isolating the genomic sequences flanking insertion elements accelerates the systematic cataloging of insertional mutants, and thus allows functions to be assigned to uncharacterized genes via reverse genetic approaches. In our current study, we report a rapid and efficient inverse PCR (iPCR) method for the isolation of gene tags in T-DNA mutant lines of rice (Oryza sativa), a model monocot plant.

Key words

Functional genomics Gene tag Inverse PCR Rice 

Notes

Acknowledgments

The methods described in this report were developed with the support of the Crop Functional Genomic Center, the 21st Century Frontier Program (Grant CG1111); from the Biogreen 21 Program, Rural Development Administration (20070401-034-001-007-03-00); from the Korea Research Foundation Grant funded by the Korean Government (MOEHRD, Basic Research Promotion Fund, KRF-2007-341-C00028); and from Kyung Hee University.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.National Research Laboratory of Plant Functional Genomics, Division of Molecular and Life Sciences, POSTECH Biotech CenterPohang University of Science and TechnologyPohangKorea
  2. 2.Graduate School of Biotechnology & Plant Metabolism Research CenterKyung Hee UniversityYonginKorea
  3. 3.Department of Plant Molecular Systems Biotechnology and Crop Biotech InstituteKyung Hee UniversityYonginRepublic of Korea

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