Replacing β-mercaptoethanol in RNA extractions

Kathleen Mommaerts*, Ignacio Sanchez, Fay Betsou, William Mathieson

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

12 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)51-53
Number of pages3
JournalAnalytical Biochemistry
Volume479
DOIs
Publication statusPublished - 15 Jun 2015

Keywords

  • DTT
  • Illustra
  • PureLink
  • RNase
  • RNeasy
  • Ribonuclease

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