Optimization of total RNA isolation from human urinary sediment

Autor(es)

Monteiro, Maria Beatriz ; Santos-Bezerra, Daniele Pereira ; Thieme, Karina ; Passarelli, Marisa ; Machado, Ubiratan Fabres ; Lin, Chin Jia ; Correa-Giannella, Maria Lucia

• Tipo de documento: Artigo Científico
• Fonte: Clinica Chimica Acta; v. 462, p. 158-161, NOV 1 2016.
• Citações Web of Science: 7

Resumo

Extracting RNA from human urinary sediment is notoriously challenging because of cell paucity and hostile environment and column-based commercial kits using silica technology are commonly used. Nonetheless, in our experience, this methodology yields low amounts of total RNA and has low rates of success. We replaced the column-based commercial kit by a protocol using guanidine isothiocyanate-phenol-chloroform buffer (Trizol reagent) followed by addition of glycogen as a carrier and precipitation with isopropanol plus sodium acetate. This methodology was more affordable and efficient for urinary sediment total RNA isolation than silica technology, resulting in higher concentrations of total RNA of better quality. (C) 2016 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 12/04831-1 - Novos moduladores do controle glicêmico e do desenvolvimento de complicações crônicas no Diabetes mellitus: perspectivas preventivas e terapêuticas
• Beneficiário: Ubiratan Fabres Machado
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 15/19000-6 - Estudo do sedimento e de micro RNAs urinários de portadores de Diabetes Tipo 1 para identificar vias associadas à progressão da doença renal diabética
• Beneficiário: Maria Beatriz Camargo Monteiro Caillaud
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado