RNA amplification for pseudogene detection using RNA-seq.
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.
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.
Stephen C M
SC
Tsoi
Michael K
MK
Dyck
D000818:Animals; D018076:DNA, Complementary; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D011544:Pseudogenes / Q000235:genetics*; D012313:RNA / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods; D013552:Swine
D018076:DNA, Complementary; D012313:RNA