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  • 1
    Publikationsdatum: 2011-08-24
    Beschreibung: Batch equilibrium adsorption isotherm determination is used to characterize the adsorption of mixed yeast RNA on agarose-immobilized m-aminophenylboronic acid. It is shown that the affinity-enhancing influence of divalent cations depends strongly on the precise nature of the cation used, with barium being far more effective than the conventionally-used magnesium. This adsorption-promoting influence of barium is suggested to arise primarily from ionic influences on the structure and rigidity of the RNA molecule, as the adsorption of ribose-based small molecules is not similarly affected. The substitution of barium for the standard magnesium counterion does not greatly promote the adsorption of DNA, implying that the effect is specific to RNA and may be useful in boronate-based RNA separations. RNA adsorption isotherms exhibit sharp transitions as functions of temperature, and these transitions occur at different temperatures with Mg2+ and with Ba2+. Adsorption affinity and capacity were found to increase markedly at lower temperatures, suggestive of an enthalpically favored interaction process. The stoichiometric displacement parameter, Z, in Ba2+ buffer is three times the value in Mg2+ buffer, and is close to unity.
    Schlagwort(e): Life Sciences (General)
    Materialart: Journal of chromatography. A; Volume 840; 2; 205-13
    Format: text
    Standort Signatur Erwartet Verfügbarkeit
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  • 2
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    Publikationsdatum: 2011-08-24
    Beschreibung: A new approach to the isolation of RNA from bacterial lysates employs selective precipitation by compaction agents, such as hexammine cobalt and spermidine. Using 3.5 mM hexammine cobalt, total RNA can be selectively precipitated from a cell lysate. At a concentration of 2 mM hexammine cobalt, rRNA can be fractionated from low molecular weight RNA. The resulting RNA mixture is readily resolved to pure 5S and mixed 16S/23S rRNA by nondenaturing anion-exchange chromatography. Using a second stage of precipitation at 8 mM hexammine cobalt, the low molecular weight RNA fraction can be isolated by precipitation. Compaction precipitation was also applied to the purification of an artificial stable RNA derived from Escherichia coli 5S rRNA and to the isolation of an Escherichia coli-expressed ribozyme. Copyright 2001 Academic Press.
    Schlagwort(e): Life Sciences (General)
    Materialart: Analytical biochemistry (ISSN 0003-2697); Volume 295; 2; 143-8
    Format: text
    Standort Signatur Erwartet Verfügbarkeit
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  • 3
    Publikationsdatum: 2011-08-24
    Beschreibung: A scaleable method for the liquid-phase separation of plasmid DNA from RNA.
    Schlagwort(e): Life Sciences (General)
    Materialart: Nature biotechnology (ISSN 1087-0156); Volume 17; 8; 822-3
    Format: text
    Standort Signatur Erwartet Verfügbarkeit
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  • 4
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    In:  Other Sources
    Publikationsdatum: 2011-08-24
    Beschreibung: The need to monitor microorganisms in the environment has increased interest in assays based on hybridization probes that target nucleic acids (e.g., rRNA). We report the development of liquid-phase assays for specific bacterial 5S rRNA sequences or similarly sized artificial RNAs (aRNAs) using molecular beacon technology. These beacons fluoresce only in the presence of specific target sequences, rendering as much as a 27-fold fluorescence enhancement. The assays can be used with both crude cell lysates and purified total RNA preparations. Minimal sample preparation (e.g., heating to promote leakage from cells) is sufficient to detect many Gram-negative bacteria. Using this approach it was possible to detect an aRNA-labeled Escherichia coli strain in the presence of a large background of an otherwise identical E. coli strain. Finally, by using a longer wavelength carboxytetramethylrhodamine beacon it was possible to reduce the fraction of the signal due to cellular autofluorescence to below 0.5%.
    Schlagwort(e): Life Sciences (General)
    Materialart: Current microbiology (ISSN 0343-8651); Volume 43; 6; 444-7
    Format: text
    Standort Signatur Erwartet Verfügbarkeit
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