Automated DNA, RNA, and Protein Extraction from Urine for Biobanking - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Journal of Biomolecular Techniques Année : 2022

Automated DNA, RNA, and Protein Extraction from Urine for Biobanking

Résumé

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 setup 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 Maxwell 16 ® system and urinary proteins were isolated by precipitation from the supernatant and the cell pellet. 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.
Fichier principal
Vignette du fichier
Article Coblance-JBT-HAL.pdf (718.91 Ko) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-04192858 , version 1 (31-08-2023)

Identifiants

Citer

Caroline Barau, Pascale Maillé, Nanor Sirab, Bijan Ghaleh, Yves Allory. Automated DNA, RNA, and Protein Extraction from Urine for Biobanking. Journal of Biomolecular Techniques, 2022, 33 (1), pp.3fc1f5fe.9e4e4f65. ⟨10.7171/3fc1f5fe.9e4e4f65⟩. ⟨hal-04192858⟩
2 Consultations
13 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More