A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis.
Anal Biochem
; 536: 8-15, 2017 11 01.
Article
em En
| MEDLINE
| ID: mdl-28803886
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.
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Base de dados:
MEDLINE
Assunto principal:
Neoplasias Ovarianas
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Análise de Sequência de RNA
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Reação em Cadeia da Polimerase Via Transcriptase Reversa
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MicroRNAs
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article