Discussion

Micro RNAs are small, naturally-occurring non-coding ribonucleic acids of approximately 22 nucleotides in length that function in gene silencing and post-transcriptional regulation of gene expression. A single microRNA (miRNA) can target and regulate several (maybe even hundreds) of transcripts, and can involve multiple biological networks or pathways. As a result, interest in miRNA biomarker research has increased. This poster describes the purification of miRNA and total RNA from fresh-frozen tissue samples using the Agencourt RNAdvance paramagnetic bead based chemistry automated on the Biomek NXP Laboratory Automated Workstation. The method enables automated purification of total RNA, including miRNA and other small RNAs, from 8–96 samples on a Biomek Span 8 workstation. Total RNA and miRNA can be purified from very small amounts of animal tissue as starting materials. Automating
SPRI (Solid Phase Reversible Immobilization) chemistry provides an easy, high yielding and robust nucleic acid
purification process that does not require centrifugation or vacuum filtration steps. Purified nucleic acids are easily eluted from the magnetic beads under aqueous conditions, which provide maximum flexibility for downstream applications. The data shows that the samples extracted using the Biomek gave comparable RNA yield, miRNA and messenger RNA gene expression compared to samples extracted manually. The RNA yield and quality is comparable to a commonly used column method using two homogenization methods.