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Polyadenylation State Microarray (PASTA) Analysis

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Yeast Systems Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 759))

Abstract

Nearly all eukaryotic mRNAs terminate in a poly(A) tail that serves important roles in mRNA utilization. In the cytoplasm, the poly(A) tail promotes both mRNA stability and translation, and these functions are frequently regulated through changes in tail length. To identify the scope of poly(A) tail length control in a transcriptome, we developed the polyadenylation state microarray (PASTA) method. It involves the purification of mRNA based on poly(A) tail length using thermal elution from poly(U) sepharose, followed by microarray analysis of the resulting fractions. In this chapter we detail our PASTA approach and describe some methods for bulk and mRNA-specific poly(A) tail length measurements of use to monitor the procedure and independently verify the microarray data.

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Acknowledgments

We thank Jürg Bähler and Daniel Lackner for their input to PASTA method development and application. This work was supported by the Victor Chang Cardiac Research Institute, the Australian Research Council, the National Health and Medical Research Council of Australia, and the Sylvia and Charles Viertel Charitable Foundation.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, Australia

    Traude H. Beilharz

  2. Genome Biology Department, The John Curtin School of Medical Research, The Australian National University, Building 131, Garran Rd, Acton (Canberra), ACT 0200, Australia

    Thomas Preiss

Authors
  1. Traude H. Beilharz
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  2. Thomas Preiss
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Correspondence to Thomas Preiss .

Editor information

Editors and Affiliations

  1. , Department of Biochemistry, University of Cambridge, Tennis Court Road 80, Cambridge, CB2 1GA, United Kingdom

    Juan I. Castrillo

  2. , Department of Biochemistry, University of Cambridge, Tennis Court Road 80, Cambridge, CB2 1GA, United Kingdom

    Stephen G. Oliver

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© 2011 Humana Press

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Beilharz, T.H., Preiss, T. (2011). Polyadenylation State Microarray (PASTA) Analysis. In: Castrillo, J., Oliver, S. (eds) Yeast Systems Biology. Methods in Molecular Biology, vol 759. Humana Press. https://doi.org/10.1007/978-1-61779-173-4_9

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  • DOI: https://doi.org/10.1007/978-1-61779-173-4_9

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-172-7

  • Online ISBN: 978-1-61779-173-4

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