Abstract
1. Fractionation of deoxribonucleic acid on paper strips or on centrifuged paper pulp.
A method is described for the fractionation of DNA preparations on DEAE-cellulose paper. This paper can be used as strips or as centrifuged paper pulp. In both cases, similar results are obtained. The reproducibility and the sensitivity of the fractionation are very high. During the fractionation process, the desorption of the macromolecular species is complete. Some of the fractions obtained can be rechromatorographed and eluted in the same position. The additivity of the fractions evidences their preexistence in the preparation. The use of cellophane strips enables the complete fractination to be run in ten hours ; with centrifuged paper pulp, a complete elution requires two hours. The physicochemical properties of the DEAE-cellulose paper allow a desorption of the highest molecular weight DNA molecules at neutral pH. These results are discussed and several examples of fractionation of polynucleotides derived from DNA are given.
2. Interrelations of the fractions
The fractions obtained by chromatography of DNA prepartaions on DEAE-cellulose paper have been characterized by different means including dialysis, alcohol and acid precipitation, optical density measurements (hyperchromic effect).
The fractionation procedure can be used to follow the effect of ageing (and of spontaneous radiolysis), ultrasonic vibrations, thermal denaturation and hydrolysis by DNAase. The concept of "DNA preparation" is discussed with respect to the specific behaviour of the different fractions into which the original mixture van be resolved.
A method is described for the fractionation of DNA preparations on DEAE-cellulose paper. This paper can be used as strips or as centrifuged paper pulp. In both cases, similar results are obtained. The reproducibility and the sensitivity of the fractionation are very high. During the fractionation process, the desorption of the macromolecular species is complete. Some of the fractions obtained can be rechromatorographed and eluted in the same position. The additivity of the fractions evidences their preexistence in the preparation. The use of cellophane strips enables the complete fractination to be run in ten hours ; with centrifuged paper pulp, a complete elution requires two hours. The physicochemical properties of the DEAE-cellulose paper allow a desorption of the highest molecular weight DNA molecules at neutral pH. These results are discussed and several examples of fractionation of polynucleotides derived from DNA are given.
2. Interrelations of the fractions
The fractions obtained by chromatography of DNA prepartaions on DEAE-cellulose paper have been characterized by different means including dialysis, alcohol and acid precipitation, optical density measurements (hyperchromic effect).
The fractionation procedure can be used to follow the effect of ageing (and of spontaneous radiolysis), ultrasonic vibrations, thermal denaturation and hydrolysis by DNAase. The concept of "DNA preparation" is discussed with respect to the specific behaviour of the different fractions into which the original mixture van be resolved.
| Original language | English |
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| Publisher | SCK CEN |
| Number of pages | 27 |
| State | Published - Dec 1964 |
Publication series
| Name | SCK CEN Reports |
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| Publisher | SCK CEN |
| No. | BLG-409 |