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 language | English (US) |
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Pages (from-to) | 420-425 |
Number of pages | 6 |
Journal | Biochemical and Biophysical Research Communications |
Volume | 366 |
Issue number | 2 |
DOIs | |
State | Published - Feb 8 2008 |
Keywords
- Adenine
- Aptamer
- IMAC
- In vitro selection
- Nickel
- Paramagnetic beads
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology