| Literature DB >> 18068116 |
Branislav Nastasijevic1, Nicole A Becker, Susan E Wurster, L James Maher.
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 (Ni(2+)-NTA) magnetic beads. Analysis of these aptamers revealed that affinity for the Ni(2+)-NTA support requires only single-stranded sequences with multiple adenosine residues. Bound nucleic acids can be eluted with imidazole. A single-stranded dA(20) affinity tag (but not other homopolymer sequences) is sufficient for immobilization of double-stranded DNA PCR products on Ni(2+)-NTA magnetic beads. Addition of an rA(20) sequence to an RNA transcript allowed its affinity capture on Ni(2+)-NTA magnetic beads, suggesting an approach for purification of poly(A) mRNA.Entities:
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Year: 2007 PMID: 18068116 DOI: 10.1016/j.bbrc.2007.11.169
Source DB: PubMed Journal: Biochem Biophys Res Commun ISSN: 0006-291X Impact factor: 3.575