RNA amplifi cation for pseudogene detection using RNA-seq

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 fi nal 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.