Replacing β-mercaptoethanol in RNA extractions.

Kathleen Mommaerts, Ignacio Sanchez, Fay Betsou, William Mathieson

Analytical biochemistry 2015 Jun 15

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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. Copyright © 2015 Elsevier Inc. All rights reserved.

Citation

Kathleen Mommaerts, Ignacio Sanchez, Fay Betsou, William Mathieson. Replacing β-mercaptoethanol in RNA extractions. Analytical biochemistry. 2015 Jun 15;479:51-3