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  • Chromatography is an integral part of large scale purification of plasmids. Samples usualy contains, except to cell components obtained after cell lysis, chromosomal DNA fragments, high molecular-weight RNA and plasmid variants. To achieve good chromatographic resolution it is necessary to degrade RNA by RNase. The aim of our work was to utilize RNase A immobilized on magnetic bead cellulose for degradation of high molecular-weight RNA before chromatographic purification of plasmid DNA. The enzyme was coupled to the particles via cyanuric chloride method. Temperature stability of soluble and immobilized RNase A was estimated after preincubation of the samples at different temperatures. Comparison of the pattern of soluble and immobilized RNase A showed a pH shift in activity. Practical use of magnetic particles in the purification of plasmid DNA is demonstrated.
  • Chromatography is an integral part of large scale purification of plasmids. Samples usualy contains, except to cell components obtained after cell lysis, chromosomal DNA fragments, high molecular-weight RNA and plasmid variants. To achieve good chromatographic resolution it is necessary to degrade RNA by RNase. The aim of our work was to utilize RNase A immobilized on magnetic bead cellulose for degradation of high molecular-weight RNA before chromatographic purification of plasmid DNA. The enzyme was coupled to the particles via cyanuric chloride method. Temperature stability of soluble and immobilized RNase A was estimated after preincubation of the samples at different temperatures. Comparison of the pattern of soluble and immobilized RNase A showed a pH shift in activity. Practical use of magnetic particles in the purification of plasmid DNA is demonstrated. (en)
Title
  • Characterisation of ribonuclease immobilised on cellulose and HEMA supports
  • Characterisation of ribonuclease immobilised on cellulose and HEMA supports (en)
skos:prefLabel
  • Characterisation of ribonuclease immobilised on cellulose and HEMA supports
  • Characterisation of ribonuclease immobilised on cellulose and HEMA supports (en)
skos:notation
  • RIV/00216224:14310/99:00003568!RIV/2002/GA0/143102/N
http://linked.open.../vavai/riv/strany
  • 69
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  • P(GA203/98/1231)
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  • 736638
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  • RIV/00216224:14310/99:00003568
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  • Characterisation of ribonuclease immobilised on cellulose and HEMA supports (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...ontrolniKodProRIV
  • [6E5E8B552A38]
http://linked.open...v/mistoKonaniAkce
  • Compiegne
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  • Compiegne
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  • 13th International Symposium of Affinity Technology and Bio-Recognition
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  • Horák, Daniel
  • Rittich, Bohuslav
  • Šafář, Jan
  • Španová, Alena
  • Lenfeld, Jiří
  • Beneš, Milan
http://linked.open...vavai/riv/typAkce
http://linked.open.../riv/zahajeniAkce
number of pages
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  • Université de Technologie de Compiegne
http://localhost/t...ganizacniJednotka
  • 14310
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