Interaction of poly(A) with different adsorbents for affinity chromatography of nucleic acids |
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Authors: | J.-L. Ochoa J. Kempf J.M. Egly |
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Affiliation: | U-184-INSERM, University Louis Pasteur, Institut de Chimie Biologique, 11, Rue Humann, Strasbourg 67085 Cedex, France |
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Abstract: | Chromatography on adsorbents for separation of mRNA containing poly(A) has given interesting results, even if the nature of the occurring interaction was not always well understood. In the present study we report the chromatographic behaviour of poly(A) homopolynucleotides on different substituted matrices: poly(U)-: poly(A)-: phenyl-, octyl-, ethanolamine-, acriflarin- and DNA-Sepharose: oligo-dT and MN-cellulose. Using different experimental conditions as ionic strength, neutral salt, pH, temperature, buffer composition it was possible to evaluate the participation of electrostatic, hydrophobic hydrogen-bonding, and/or charge-transfer interaction. Furthermore, it is shown that poly(A) interacts non-specifically with matrices like acriflavin or DNA-Sepharose, as well as with oligo-dT cellulose or poly(U)-Sepharose. |
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