The Determination of Nucleic Acids

Munro, Hamish N.; Fleck, A.

doi:10.1002/9780470110324.ch5

Cited by 853 publications

(57 citation statements)

References 302 publications

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“…These were exactly the changes in nucleic acids exhibited by follicle cells during in situ termination of vitellogenesis (Wang and Telfer, 2000). In the present work, the effect of chlorfluazuron on the DNA and RNA contents was estimated according to the method described by Burton (1956) and Munro (1966). The general growth of the insects is controlled by moulting hormone (MH) and juvenile hormone (JH) whose metabolism and degradation are checked by enzymes.…”

Section: Discussionmentioning

confidence: 99%

Biochemical Analyses of Action of Chlorfluazuron as Reproductive Inhibitor in Spodoptera litura

Perveen¹

2012

Insecticides - Advances in Integrated Pest Management

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Section: Discussionmentioning

confidence: 99%

Biochemical Analyses of Action of Chlorfluazuron as Reproductive Inhibitor in Spodoptera litura

Perveen¹

2012

Insecticides - Advances in Integrated Pest Management

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“…Although nucleic acids can be measured spectrophotometrically after lengthy extraction and purification (Schmidt and Tannhauser, 1945;Munro and Fleck, 1966), the procedure lacks the sensitivity needed for small biomasses associated with microplanktonic samples. Increased sensitivity can be achieved using fluorochromes that specifically bind to DNA or that bind to both DNA and RNA, without previous purification or separation of the two molecules.…”

Section: Introductionmentioning

confidence: 99%

Quantifying RNA and DNA in planktonic organisms with SYBR Green II and nucleases. Part A. Optimisation of the assay

Berdalet¹,

Roldán²,

Olivar³

et al. 2005

Sci. Mar.

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SUMMARY: Assay protocols for RNA and DNA in crude plankton extracts using the fluorochrome SYBR Green II are developed here. The method is based on the fluorescence in 3 aliquots: the first measures RNA after DNA digestion; the second measures DNA after RNA digestion; and the third measures residual fluorescence after digestion of both DNA and RNA. This residual fluorescence measurement is critical for accurate calculations of the nucleic acids. Optimisation of the assay conditions are described: fluorochrome concentration, buffer composition, fluorescence stability, temperature and duration of nuclease incubation. In the optimised procedure, the assays are performed in 5 mM Tris buffer (containing 0.9 mM CaCl 2 ·2H 2 O and 0.9 mM MgCl 2 ·6H 2 O, pH 8.0); DNase and RNase incubations are conducted at 37ºC for 20 min; the fluorochrome is added to all assays at a final concentration of 3.5x10 -4 and readings are done within the 10-60 min period following the SYBR Green II addition. The study evidenced the importance of the residual fluorescence after nuclease digestion, which is especially taken into account in the calculation of the nucleic acid concentrations. Finally, the variability of the fluorescent response to different RNA and DNA standards is examined; from the performed tests, calculations are based on rRNA from calf liver and DNA from calf thymus standards. The accompanying paper (Berdalet et al., 2005) describes the development of the extraction protocol, as well as the application of both protocols in measuring RNA/DNA ratios in natural plankton samples, and a comparison with ethidium bromide based methods.Key words: SYBR Green II, DNase, RNase, RNA/DNA ratios, plankton. RESUMEN: CUANTIFICACIÓN DE ARN Y ADN EN ORGANISMOS PLANCTÓNICOS MARINOS MEDIANTE SYBR GREEN II Y NUCLE-ASES. PARTE A. OPTIMIZACIÓN DEL ENSAYO. -En este trabajo se desarrollan los protocolos para la cuantificación de ARN y ADN en extractos no purificados de plancton utilizando SYBR Green II. El método se basa en la fluorescencia de 3 alícuo-tas: la primera mide el ARN tras la digestión del ADN; la segunda mide el ADN tras la digestión del ARN y la tercera mide la fluorescencia residual tras la digestión tanto del ARN como del ADN. La medida de esta fluorescencia residual es críti-ca para obtener una buena estimación de los ácidos nucleicos. Se describen las condiciones de optimización del ensayo: concentración de fluorocromo, composición del tampón, estabilidad de la fluorescencia, temperatura y duración de la incubación con nucleasas. En el procedimiento optimizado los ensayos se realizan en tampón Tris 5 mM (0.9 mM CaCl 2 ·2H 2 O y 0.9 mM MgCl 2 ·6H 2 O, pH 8); las incubaciones con nucleasas se llevan a cabo a 37°C durante 20 min; el fluorocromo se añade a todos los ensayos a una concentración final de 3.5X10 -4 y las lecturas se realizan en los 10-60 min posteriores a la adición de SYBR Green II. Este estudio evidencia la importancia de la fluorescencia residual después de la digestión con nucleasas, la cual es especialmente incluída...

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“…The concentration of protein in the gastrocnemius muscle, liver, and brain was determined according to the method described by Lowry et al (1951), while the concentration of RNA in these tissues was determined according to the method described by Munro and Fleck (1966). The concentration of DNA was obtained by the method described by Gendimenico, Bouquin and Tramposch (1988).…”

Section: Biochemical Analysismentioning

confidence: 99%

Early weaning impairs body composition in male mice

Borges

Rogero

Pires

et al. 2009

Braz. J. Pharm. Sci.

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This study aimed to evaluate the effect of early weaning on body composition and on parameters related to nutritional status in mice. The experimental group consisted of male Swiss Webster mice that were weaned early (at postnatal day fourteen) and fed an appropriate diet for growing rodents until postnatal day twenty-one (EW group). The control group consisted of male mice breastfed until postnatal day twenty-one (CON group). All animals were sacrificed on the twenty-first day of life. The EW group showed a decrease in liver and muscle protein content and concentration, brain protein concentration, brain DNA content and concentration, as well as liver and muscle protein/RNA ratio (p<0.05). Concerning body composition, the EW mice showed increased moisture content, increased moisture and lipid percentage, and a smaller percentage and content of protein and ash in the carcass (p<0.05). These results indicate that early weaning impairs body composition and parameters related to nutritional status, which may be explained by retarded chemical maturation processes. This data may contribute to the overall understanding of the influence of breastfeeding versus feeding with artificial milk on body composition and on nutritional status. Uniterms:Weaning. Body composition. Breastfeeding. Nutritional status.O presente estudo objetivou avaliar o efeito do desmame precoce sobre a composição corporal e sobre parâmetros indicativos do estado nutricional de camundongos. O grupo experimental consistiu de camundongos Swiss Webster, machos, desmamados precocemente (14º dia de vida) e alimentados com ração apropriada para roedores em crescimento até o 21º dia pós-natal (grupo DESM). O grupo controle consistiu de camundongos amamentados até o 21º dia pós-natal (grupo CON). Todos os animais foram sacrificados no 21º dia de vida. O grupo DESM apresentou redução da concentração e conteúdo hepático e muscular de proteínas, da concentração cerebral de proteínas, da concentração e conteúdo cerebral de DNA e da razão proteína/RNA hepática e muscular (p<0,05). Quanto à composição corporal, o grupo DESM apresentou maior conteúdo de umidade, maior percentual de umidade e lipídios e menor conteúdo e percentual de cinzas e proteína na carcaça (p<0,05). Os resultados indicam que o desmame precoce acarreta em prejuízo à composição corporal e a parâmetros indicativos do estado nutricional, o que pode estar relacionado ao retardo do processo de maturação química. Os dados do presente estudo podem contribuir para o entendimento da influência da alimentação com fórmulas infantis sobre a composição corporal e sobre o estado nutricional.Unitermos: Desmame. Composição corporal. Aleitamento materno. Estado nutricional.

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The Determination of Nucleic Acids

Cited by 853 publications

References 302 publications

Biochemical Analyses of Action of Chlorfluazuron as Reproductive Inhibitor in Spodoptera litura

Biochemical Analyses of Action of Chlorfluazuron as Reproductive Inhibitor in Spodoptera litura

Quantifying RNA and DNA in planktonic organisms with SYBR Green II and nucleases. Part A. Optimisation of the assay

Early weaning impairs body composition in male mice

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