Urinary microvesicle RNA integrity and alignment to the genome.

<p>a) RNA isolated from urinary microvesicles was shown to be of high integrity with prominent 18S and 28S rRNA peaks when analyzed using the Agilent Bioanalyzer. Red trace 1.7 ng RNA with DNase, Blue trace 2.2 ng RNA without DNase. b) Flow chart outlining sample processing. An initial RNase and DNase digestion was carried out to remove extraneous nucleic acids co-isolating with the microvesicle pellet. To determine the proportion of potential DNA inside the microvesicles the extracted RNA was divided into two groups; No DNase digestion (-DNase), which yields RNA+DNA and DNase digested (+DNase) which yields RNA. c) Both the -DNase and the +DNase samples showed a similar trend in read distribution with ∼88% of reads mapping to rRNA, ∼4% mapping to genes and ∼6% mapping to ncRNA. A smaller proportion of reads (<0.1%) mapped to mitochondrial genome. Approximately 2% of reads failed to hit the human genome. (mito – mitochondrial).