Abstract
Transcriptomic research using microarrays and RNA-Sequencing (RNA-seq) is now possible starting from minute biological samples, such as clinical specimens or embryos, due to the development of highly sensitive and reproducible cDNA synthesis methods. Here, we describe a quick method of RNA amplification and double-stranded cDNA synthesis starting with 10 ng of high-quality total RNA extracted from porcine embryos. The final product (double-stranded DNA) is adequate for the detection by RNA-seq of protein-coding transcripts, as well as of all the other classes of noncoding RNAs, including pseudogenes.
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Acknowledgements
This research was supported by the Natural Science and Engineering Research Council of Canada, EmbryoGENE Strategic Research Network.
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© 2014 Springer Science+Business Media New York
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Tsoi, S.C.M., Dyck, M.K. (2014). RNA Amplification for Pseudogene Detection Using RNA-Seq. In: Poliseno, L. (eds) Pseudogenes. Methods in Molecular Biology, vol 1167. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0835-6_9
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DOI: https://doi.org/10.1007/978-1-4939-0835-6_9
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-0835-6
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