Abstract
Sequencing-based whole-transcriptome analysis (i.e., RNA-Seq) can be a powerful tool when used to measure gene expression, detect novel transcripts, characterize transcript isoforms, and identify sequence polymorphisms. However, this method can be inefficient when the goal is to study only one component of the transcriptome, such as long noncoding RNAs (lncRNAs), which constitute only a small fraction of transcripts in a total RNA sample. Here, we describe a target enrichment method where a total RNA sample is converted to a sequencing-ready cDNA library and hybridized to a complex pool of lncRNA-specific biotinylated long oligonucleotide capture probes prior to sequencing. The resulting sequence data are highly enriched for the targets of interest, dramatically increasing the efficiency of next-generation sequencing approaches for the analysis of lncRNAs.
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Acknowledgement
NIMBLEGEN and SEQCAP are trademarks of Roche.
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For life science research only. Not for use in diagnostic procedures.
© 2015 Roche NimbleGen, Inc.
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© 2016 Springer Science+Business Media New York
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Tan, J.C., Bouriakov, V.D., Feng, L., Richmond, T.A., Burgess, D. (2016). Targeted LncRNA Sequencing with the SeqCap RNA Enrichment System. In: Feng, Y., Zhang, L. (eds) Long Non-Coding RNAs. Methods in Molecular Biology, vol 1402. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3378-5_8
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DOI: https://doi.org/10.1007/978-1-4939-3378-5_8
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3376-1
Online ISBN: 978-1-4939-3378-5
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