Literature DB >> 27666760

Optimization of total RNA isolation from human urinary sediment.

Maria Beatriz Monteiro1, Daniele Pereira Santos-Bezerra1, Karina Thieme1, Marisa Passarelli2, Ubiratan Fabres Machado3, Chin Jia Lin4, Maria Lúcia Corrêa-Giannella5.   

Abstract

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.
Copyright © 2016 Elsevier B.V. All rights reserved.

Entities:  

Keywords:  Chronic kidney disease; Gene expression; RNA; RNA isolation; Urinary sediment

Mesh:

Substances:

Year:  2016        PMID: 27666760     DOI: 10.1016/j.cca.2016.09.018

Source DB:  PubMed          Journal:  Clin Chim Acta        ISSN: 0009-8981            Impact factor:   3.786


  8 in total

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  8 in total

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