title: Replacing β-mercaptoethanol in RNA extractions.
abstract: RNA extractions are potentially compromised in terms of both yield and quality by ribonucleases (RNases). The pungent and toxic reducing agent β-mercaptoethanol (β-ME), therefore, is commonly added to the biospecimen's lysis buffer to aid in RNase deactivation. Using different tissue types (liver tissue, kidney tissue, and cell pellets), extraction kits (RNeasy Mini Kit, Illustra RNA Spin Mini Kit, and PureLink Mini Kit), RNA quality assays (RNA integrity numbers [RINs] and quantitative real-time polymerase chain reaction [qRT-PCR]), yield assessments, and in vitro functional RNase assays (RNaseAlert Kit), we demonstrate that β-ME should be replaced by the less toxic dithiothreitol (DTT) alternative.
authors.affiliation: Integrated Biobank of Luxembourg, L-1210 Luxembourg, Luxembourg. Electronic address: kathleen.mommaerts@ibbl.lu.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Mommaerts
authors.affiliation: Integrated Biobank of Luxembourg, L-1210 Luxembourg, Luxembourg.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Sanchez
authors.affiliation: Integrated Biobank of Luxembourg, L-1210 Luxembourg, Luxembourg.
authors.forename: Fay
authors.initials: F
authors.lastname: Betsou
authors.affiliation: Integrated Biobank of Luxembourg, L-1210 Luxembourg, Luxembourg.
authors.forename: William
authors.initials: W
authors.lastname: Mathieson
mesh_terms: D000818:Animals; D004229:Dithiothreitol / Q000737:chemistry*; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008623:Mercaptoethanol / Q000737:chemistry*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification*; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
chemical_list: D008623:Mercaptoethanol; D012313:RNA; D004229:Dithiothreitol
keywords: DTT; Illustra; PureLink; RNase; RNeasy; Ribonuclease