A recent study by Anson Ku and others from Lund University, “A urinary extracellular vesicle microRNA biomarker discovery pipeline; from automated extracellular vesicle enrichment by acoustic trapping to microRNA sequencing”, investigated a comprehensive biomarker discovery pipeline for miRNA derived from both small and large urinary EVs.
Biomarker Discovery Pipeline
The biomarker discover pipeline consists of:
- Enrichment of urinary EVs using an acoustic trap or ultracentrifugation
- RNase A treatment to eliminate contaminating RNA molecules
- Total RNA isolation
- Small RNA library preparation
- Illumina® sequencing
Low Input miRNA Library Prep
Libraries were constructed from two commercially available, low-input compatible, small RNA library kits, the NEXTFLEX® Small RNA-seq kit and the CATS™ small RNA library (Diagenode, USA), using an estimated 130 pg of input total RNA isolated form 1.7 mL of urine. The NEXTFLEX® Small RNA-seq kit because the NEXTFLEX® libraries resulted in more mappable reads compared to the Diagenode® CATS™ libraries which had a predominant fraction of rRNA reads. Additionally, the NEXTFLEX® Small RNA-seq kit was the only kit able to successfully generate libraries from all the low-input samples.
Read the research:
Ku, A., Ravi, N., Yang, M., Evander, M., Laurell, T., Lilja, H., & Ceder, Y. (2019). A urinary extracellular vesicle microRNA biomarker discovery pipeline; from automated extracellular vesicle enrichment by acoustic trapping to microRNA sequencing. Plos One, 14(10). doi: 10.1371/journal.pone.0224604.
Learn more about the NEXTFLEX® Small RNA-seq kit and improve your miRNA biomarker pipeline.