Sequence-specific binding of DNA and RNA to immobilized Nickel ions

Branislav Nastasijevic, Nicole A. Becker, Susan E. Wurster, L. James Maher

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Immobilization of divalent Nickel cations provides a tool for affinity purification of proteins containing hexahistidine tags. During experiments to generate single-stranded DNA aptamers to immobilized proteins we inadvertently identified DNA sequences with affinity for Nickel-nitrilotriacetic acid (Ni2+-NTA) magnetic beads. Analysis of these aptamers revealed that affinity for the Ni2+-NTA support requires only single-stranded sequences with multiple adenosine residues. Bound nucleic acids can be eluted with imidazole. A single-stranded dA20 affinity tag (but not other homopolymer sequences) is sufficient for immobilization of double-stranded DNA PCR products on Ni2+-NTA magnetic beads. Addition of an rA20 sequence to an RNA transcript allowed its affinity capture on Ni2+-NTA magnetic beads, suggesting an approach for purification of poly(A) mRNA.

Original languageEnglish (US)
Pages (from-to)420-425
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume366
Issue number2
DOIs
StatePublished - Feb 8 2008

Keywords

  • Adenine
  • Aptamer
  • IMAC
  • In vitro selection
  • Nickel
  • Paramagnetic beads

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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