A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis

Kun Zhou, Monique A. Spillman, Kian Behbakht, Julia M. Komatsu, Juan E Abrahante - Llorens, Douglas Hicks, Brent Schotl, Evan Odean, Kenneth L. Jones, Michael W. Graner, Lynne T Bemis

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Readily accessible samples such as urine or blood are seemingly ideal for differentiating and stratifying patients; however, it has proven a daunting task to identify reliable biomarkers in such samples. Noncoding RNA holds great promise as a source of biomarkers distinguishing physiologic wellbeing or illness. Current methods to isolate and characterize RNA molecules in urine are limited. In this proof of concept study, we present a method to extract and identify small noncoding RNAs in urine. Initially, quantitative reverse transcription PCR was applied to confirm the presence of microRNAs in total RNA extracted from urine. Once the presence of micro RNA in urine was confirmed, we developed a method to scale up RNA extraction to provide adequate amounts of RNA for next generation sequence analysis. The method described in this study is applicable to detecting a broad range of small noncoding RNAs in urine; thus, they have wide applicability for health and disease analyses.

Original languageEnglish (US)
Pages (from-to)8-15
Number of pages8
JournalAnalytical Biochemistry
Volume536
DOIs
StatePublished - Nov 1 2017

Fingerprint

Urine
RNA
Small Untranslated RNA
Polymerase Chain Reaction
Biomarkers
MicroRNAs
Untranslated RNA
Transcription
Blood
Health
Reverse Transcription
Molecules
Sequence Analysis

Keywords

  • Extracellular RNA in urine
  • Noncoding RNA
  • Ovarian cancer
  • Small RNA next generation sequencing
  • tRNA fragments

Cite this

A method for extracting and characterizing RNA from urine : For downstream PCR and RNAseq analysis. / Zhou, Kun; Spillman, Monique A.; Behbakht, Kian; Komatsu, Julia M.; Abrahante - Llorens, Juan E; Hicks, Douglas; Schotl, Brent; Odean, Evan; Jones, Kenneth L.; Graner, Michael W.; Bemis, Lynne T.

In: Analytical Biochemistry, Vol. 536, 01.11.2017, p. 8-15.

Research output: Contribution to journalArticle

Zhou, K, Spillman, MA, Behbakht, K, Komatsu, JM, Abrahante - Llorens, JE, Hicks, D, Schotl, B, Odean, E, Jones, KL, Graner, MW & Bemis, LT 2017, 'A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis', Analytical Biochemistry, vol. 536, pp. 8-15. https://doi.org/10.1016/j.ab.2017.08.003
Zhou, Kun ; Spillman, Monique A. ; Behbakht, Kian ; Komatsu, Julia M. ; Abrahante - Llorens, Juan E ; Hicks, Douglas ; Schotl, Brent ; Odean, Evan ; Jones, Kenneth L. ; Graner, Michael W. ; Bemis, Lynne T. / A method for extracting and characterizing RNA from urine : For downstream PCR and RNAseq analysis. In: Analytical Biochemistry. 2017 ; Vol. 536. pp. 8-15.
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