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Methods Mol Biol. 2014;1167:119-27. doi: 10.1007/978-1-4939-0835-6_9.

RNA amplification for pseudogene detection using RNA-seq.

Author information

1
Agricultural Genomics and Proteomics Unit, Core Research Laboratory, Department of Agricultural, Food and Nutritional Science, 4-32G Agriculture/Forestry Centre, University of Alberta, Edmonton, AB, Canada, T6G 2P5, scmtsoi@gmail.com.

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.

PMID:
24823775
DOI:
10.1007/978-1-4939-0835-6_9
[Indexed for MEDLINE]

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