Optimization of small RNA library preparation protocol from human urinary exosomes.

ABSTRACT:

Background

Sequencing of miRNAs isolated from exosomes has great potential to identify novel disease biomarkers, but exosomes have low amount of RNA, hindering adequate analysis and quantification. Here, we have assessed several steps in developing an optimized small RNA (sRNA) library preparation protocol for next-generation sequencing (NGS) miRNA analysis from urinary exosomes.

Methods

A total of 24 urinary exosome samples from donors were included in this study. RNA was extracted by column-based methods. The quality of extracted RNA was assessed by spectrophotometric quantification and Bioanalyzer software analysis. All libraries were prepared using the CleanTag small RNA library preparation protocol and the effect of our additional modifications on adapter-dimer presence

SUBMITTER: Olivares D 

PROVIDER: S-EPMC7081560 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature