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Strand-Specific RNA-seq Applied to Malaria Samples

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RNA Abundance Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 883))

Abstract

Over the past few years only, next-generation sequencing technologies became accessible and many applications were rapidly derived, such as the development of RNA-seq, a technique that uses deep sequencing to profile whole transcriptomes. RNA-seq has the power to discover new transcripts and splicing variants, single-nucleotide variations, fusion genes, and mRNA level-based expression profiles. Preparing RNA-seq libraries can be delicate and usually obligates buying expensive kits that require large amounts of stating materials. The method presented here is flexible and cost-effective. Using this method, we prepared high-quality strand-specific RNA-seq libraries from RNA extracted from the human malaria parasite Plasmodium falciparum. The libraries are compatible with Illumina®’s sequencers Genome Analyzer and Hi-Seq. The method can, however, be easily adapted to other platforms.

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References

  1. Le Roch KG et al (2003) Discovery of gene function by expression profiling of the malaria parasite life cycle. Science 301(5639):1503–1508

    Article  PubMed  Google Scholar 

  2. Bozdech Z et al (2003) The transcriptome of the intraerythrocytic developmental cycle of Plasmodium falciparum. PLoS Biol 1(1):E5

    Article  PubMed  Google Scholar 

  3. Otto TD et al (2010) New insights into the blood-stage transcriptome of Plasmodium ­falciparum using RNA-Seq. Mol Microbiol 76(1):12–24

    Article  PubMed  CAS  Google Scholar 

  4. Katherine Sorber MT, Dimon JLDeRisi (2011) RNA-Seq analysis of splicing in Plasmodium falciparum uncovers new splice junctions, alternative splicing and splicing of antisense transcripts. Nucleic Acids Res 39(9): 3820–3835

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Acknowledgments

The authors thank Courtney Brady (NuGEN®), and Barbara Walter, John Weger, Rebecca Sun, and Glenn Hicks (Institute for Integrative Genome Biology, University of California Riverside) for their assistance in the library preparation and sequencing processes.

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Correspondence to Karine G. Le Roch .

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© 2012 Springer Science+Business Media, LLC

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Ponts, N., Chung, DW.D., Le Roch, K.G. (2012). Strand-Specific RNA-seq Applied to Malaria Samples. In: Jin, H., Gassmann, W. (eds) RNA Abundance Analysis. Methods in Molecular Biology, vol 883. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-839-9_4

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  • DOI: https://doi.org/10.1007/978-1-61779-839-9_4

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-838-2

  • Online ISBN: 978-1-61779-839-9

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