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


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.

SUBMITTER: Zhou K 

PROVIDER: S-EPMC6278937 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature

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

Zhou Kun K   Spillman Monique A MA   Behbakht Kian K   Komatsu Julia M JM   Abrahante Juan E JE   Hicks Douglas D   Schotl Brent B   Odean Evan E   Jones Kenneth L KL   Graner Michael W MW   Bemis Lynne T LT  

Analytical biochemistry 20170810


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.  ...[more]

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