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
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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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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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