Abstract
Various methods for extracting nucleic acids from pollen were tested to find a suitable procedure for obtaining a pure preparation of nucleic acids uncontaminated by polysaccharides and polyphosphates without the use of ion exchangers. Extraction was carried out with perchloric acid, potassium hydroxide, ribonuclease and deoxyribonuclease, sodium tetraborate, and combinations of these. In all fractions, residues of precipitates and residues of extracted pollen matter, the quantity of RNA, DNA, proteins and concomitants,i.e. Polysaccharides and polyphosphates, was determined. The purity of preparations was checked by means of UV-spectra. The criterion of nucleic acid purity was agreement between the nucleic acid amounts calculated on the basis of measurement of absorption in UV-region, orcinol reaction and content of phosphorus. It was found that in our material only a few methods would be applicable and with great limitation, because many polysaccharides and polyphosphates appeared in nucleic acid fractions.
Abstract
Experimentálně byly vyzkoušeny různé metody extrakoe nukleových kyselin z pylového materiálu za účelem najít dobrý postup pro získání čistého preparátu nukleových kyselin nekontaminovaného polysacharidy a polyfosforečnany bez užití iontoměničů: Extrakce s lO4, KOH, ribonukleásou a desoxyribonukleásou, Na2B4O7 a jejich kombinacemi. U všech frakcí, zbytků sraženin a extrahované pylové hmoty byl stanoven obsah BNK, DNK, bílkovin a kontamiaujících látek, tj. polysacharidů a polyfosforečnanů. Čistota preparátu byla kontrolována sledováním průběhu UV-spekter. Za kritérium čistoty byla považována shoda mezi množatvím nukleových kyselin měřeném v UV-oblasti, orcinolovou metodou a přes fosfor. Bylo nalezeno, žpro náš pokusný materiál lze užít pouze několik metod extrakce a to ještě velmi omezeně nebot ve frakcích nukleových kyselin bylo nalezeno velké množství polysacharidů a polyfosfo-rečnanů.
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Süss, J. Use of acid, alkali and enzymes for the extraction of nucleic acids from pollen. Biol Plant 12, 332–341 (1970). https://doi.org/10.1007/BF02922187
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DOI: https://doi.org/10.1007/BF02922187