Efficient cellular fractionation improves RNA sequencing analysis of mature and nascent transcripts from human tissues.

ABSTRACT:

Background

The starting material for RNA sequencing (RNA-seq) studies is usually total RNA or polyA+ RNA. Both forms of RNA represent heterogeneous pools of RNA molecules at different levels of maturation and processing. Such heterogeneity, in addition to the biases associated with polyA+ purification steps, may influence the analysis, sensitivity and the interpretation of RNA-seq data. We hypothesize that subcellular fractions of RNA may provide a more accurate picture of gene expression.

Results

We present results for sequencing of cytoplasmic and nuclear RNA after cellular fractionation of tissue samples. In comparison with conventional polyA+ RNA, the cytoplasmic RNA contains a significantly higher fraction of exonic sequence, providing increased sensitivity in expressio

SUBMITTER: Zaghlool A 

PROVIDER: S-EPMC3833653 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature