Separation of phenol and ribonucleic acid by dextran gel filtration (sephadex G-25)

Gross, Maria; Skoczylas, Bogna; Turski, W.

doi:10.1016/0003-2697(65)90036-9

Cited by 7 publications

(3 citation statements)

References 7 publications

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“…The mixture was shaken for 15min and centrifuged at 10000g for lOn1in. Th's treatmIent was repeated until the complete disappearance of the interphase; this step is a modification of the coldphenol procedure described by Cartouzou et al (1968), introduced to complete the deproteinization (Sevag et al, 1938) and also to remove the phenol (Gross et al, 1965).…”

Section: Methodsmentioning

confidence: 99%

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Synthesis of ribonucleic acid in the venom gland of Crotalus durissus terrificus (Ophidia, Reptilia) after manual extraction of the venom

Lucca¹,

Imaizumi²

1972

Biochemical Journal

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1. The incorporation of [5-3H]uridine into RNA of the venom gland of Crotalus durissus terrificus was studied after manual extraction (‘milking’) of the venom. The labelled precursor was injected immediately after milking. 2. The RNA was extracted 1, 2, 4, 6 and 8h after injection of the label and analysed by sucrose-density-gradient centrifugation. 3. The sedimentation analysis showed that 18S rRNA synthesis is higher than 28S rRNA at all time-intervals. The specific radioactivities of both ribosomal components did not reach a plateau even at 8h after injection. 4. An RNA fraction was detected sedimenting between 18S rRNA and 4S tRNA and was called the 10–14S fraction. The specific radioactivity was always higher than that of both classes of rRNA and reached the maximum value at 6h of labelling. 5. The incorporation of the precursor was also studied by radioautography, which helped to elucidate the intracellular origin of the RNA analysed by sucrose-density-gradient centrifugation.

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Section: Methodsmentioning

confidence: 99%

“…The yield was 3.7mg/g of fresh venom gland. The possibility of contamination by protein and phenol was checked by examination of the E260/E280 ratio (Rousseau & Crabbe, 1968) and the E220/E260 ratio (Gross et al, 1965) respectively. The test described by Burton (1956) indicated a negligible contamination by DNA.…”

Section: Methodsmentioning

confidence: 99%

Synthesis of ribonucleic acid in the venom gland of Crotalus durissus terrificus (Ophidia, Reptilia) after manual extraction of the venom

Lucca¹,

Imaizumi²

1972

Biochemical Journal

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“…The use of Sephadex gel in the preparation of RNA fractions was noted previously (Gross, Skoczylas & Turski, 1965;McCoy & Carter, 1968); however, its application to removal of phenol in the preparation of tRNA appears to have been neglected. The high affinity of Sephadex for phenol (Determann & Walter, 1968) no doubt accounts for the effective separation of large amounts of RNA from phenol that can readily be accomplished by this procedure.…”

Section: Discussionmentioning

confidence: 99%

Extensive charging of transfer ribonucleic acid by bean leaf extracts in vitro

Tao

Hall

1971

Biochemical Journal

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1. Phenol was effectively removed from aqueous extracts of RNA by chromatography on Sephadex G-50. 2. Elution of tRNA from Sephadex G-50 columns at pH7.6 was shown to remove 91% of the endogenously bound amino acids. 3. tRNA prepared without recourse to ethanolic precipitation was capable of accepting much greater amounts of amino acids than could redissolved samples of precipitated tRNA. 4. Aminoacyl-tRNA synthetase enzymes were partially purified with calcium phosphate gel. Elution of enzymes from the gel at pH6.5 yielded a fraction having phenylalanine- and alanine-charging activity, but no aspartate-, lysine- or proline-charging activity, whereas elution at pH7.6 gave a fraction having aspartate-, lysine- and proline-charging activity but no phenylalanine- or alanine-charging activity. 5. By using partially synthetase enzymes and tRNA eluted from DEAE-Sephadex A-50 columns, 52% of the theoretical maximum of aminoacyl-tRNA synthesis was obtained in vitro.

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Determann

1967

Gelchromatographie

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Separation of phenol and ribonucleic acid by dextran gel filtration (sephadex G-25)

Cited by 7 publications

References 7 publications

Synthesis of ribonucleic acid in the venom gland of Crotalus durissus terrificus (Ophidia, Reptilia) after manual extraction of the venom

Synthesis of ribonucleic acid in the venom gland of Crotalus durissus terrificus (Ophidia, Reptilia) after manual extraction of the venom

Extensive charging of transfer ribonucleic acid by bean leaf extracts in vitro

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