Human Urinary Rna Exosome: Optimizing Methods Of Isolations And Sample Preparation For Transcriptome

  • R. Danarto Universitas Gadjah Mada
  • Angga Dwi Prasetyo
  • Joni Kristanto
  • Richardus Hugo
  • Santosa Pradana Putra Setya Negara
  • Tirta Wardana
  • Indwiani Astuti
  • Sofia Mubarika Haryana
Keywords: urine, lysis buffer, RNA isolation, stability


Introduction: Human urine is potential bio-fluid to study as a diagnostic biomarker methods. There are substances that secreted from metabolism residue and damaged cell, including genetic substances that cast away through urine, and also RNA (Ribonucleic Acid). Recently, RNA (coding-ncRNAs) has been developed for diagnosis method because it could represent profile expression in the cell. Furthermore, RNA urinary isolation methods is  important to be established to get reliable biomarker non-invasive compared to tissue biopsy. In fact that optimization method for isolation RNA from urine is not clear. So that,  the optimization and stability storage study are needed to be used as reference standard protocol. This study performed  to know  the optimization methods of RNA isolation from urine samples and RNA concentration stability storage for transcriptomic (non-coding RNA) analysis. Methods: Each sample was collected as many 15 mL in the morning and treated with lysis solution from different manufacturers (Qiagen, Ambion, Geneid, control without buffer). ANOVA Statistic analysis was performed to know significant difference between methods used. Results: RNA stability measuring of RNA and DNA observed on day 1, 3, 5, 7, 9, 11, and 13-day by P-value > 0.01. at the same time, RNA stability storage is known to decrease consistently by 0.1-1 ng each day. Quantifikasi miRNA could be done from urine samples. Conclusion: There is no significant difference between all the methods used.