Your browser doesn't support javascript.
loading
Automated DNA, RNA, and Protein Extraction from Urine for Biobanking.
Barau, Caroline; Maillé, Pascale; Sirab, Nanor; Ghaleh, Bijan; Allory, Yves.
Afiliação
  • Barau C; APHP, Hôpital Henri Mondor, Plateforme de Ressources Biologiques CréteilF-94000 France.
  • Maillé P; APHP, Hôpital Henri Mondor, Département de Pathologie CréteilF-94000 France.
  • Sirab N; APHP, Hôpital Henri Mondor, Département de Pathologie CréteilF-94000 France.
  • Ghaleh B; APHP, Hôpital Henri Mondor, Plateforme de Ressources Biologiques CréteilF-94000 France.
  • Allory Y; Institut Curie, CNRS, UMR 144 Paris75248 France.
J Biomol Tech ; 33(1)2022 04 15.
Article em En | MEDLINE | ID: mdl-35836999
Introduction/Objective: DNA, RNA, and proteins are unavoidable human biomarkers. Today, blood remains the commonly used source of biomarkers despite numerous limitations. Therefore, other sources of biomarkers such as urine could be more appropriate for research in the field of bladder cancer. The aim of this study was to set up a new automated procedure for urinary DNA, RNA, and protein extraction and to evaluate their quality and quantity. Materials and Methods: This study was conducted in the setting of the COBLAnCE cohort. Urinary DNA and RNA were extracted using the Maxwell 16 system, and urinary proteins were isolated by precipitation from the supernatant and the cell pellet. The concentration and purity of nucleic acids were determined by spectrophotometry. RNA integrity was determined by the Agilent Bioanalyzer. PCR assays were also used to ensure the quality of DNA and RNA samples. The quality of protein samples obtained was determined by Western blot analysis. Results: PCR experiments performed highlighted that it is possible to use the DNA and RNA samples for amplification, gene expression, or genotyping. However, DNA and RNA recovery from urine was highly variable among patients, with a significant impact of the patient's gender. The samples were highly degraded. Finally, our protocol of protein isolation was effective in extracting urinary supernatant proteins as well as pellet proteins. Discussion: Therefore, urine samples could constitute valuable resources for subsequent investigations in bladder cancer. These samples will allow identifying new easy-access biomarkers for the early detection of cancer, monitoring cancer progression, and assessing response to therapy.
Assuntos
Palavras-chave

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Neoplasias da Bexiga Urinária / RNA Tipo de estudo: Diagnostic_studies / Guideline / Prognostic_studies / Screening_studies Limite: Humans Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Neoplasias da Bexiga Urinária / RNA Tipo de estudo: Diagnostic_studies / Guideline / Prognostic_studies / Screening_studies Limite: Humans Idioma: En Ano de publicação: 2022 Tipo de documento: Article