Nuclear RNA Isolation and Sequencing

Most transcriptome studies involve sequencing and quantification of steady-state mRNA by isolating and sequencing poly (A) RNA. Although this type of sequencing data is informative to determine steady-state mRNA levels it does not provide information on transcriptional output and thus may not always reflect changes in transcriptional regulation of gene expression. Furthermore, sequencing poly (A) RNA may miss transcribed regions of the genome not usually modified by polyadenylation which includes many long noncoding RNAs. Here, researchers from the University of Toronto describe nuclear-RNA sequencing (nucRNA-seq) which investigates the transcriptional landscape through sequencing and quantification of nuclear RNAs which are both unspliced and spliced transcripts for protein-coding genes and nuclear-retained long noncoding RNAs.


Relative quantities of each sequence in the nuclear (NucRNA) and cytoplasmic (CytoRNA) fractions

Dhaliwal NK, Mitchell JA. (2016) Nuclear RNA Isolation and Sequencing. Methods Mol Biol 1402:63-71. [abstract]

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