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At least 505 records · Page 28Linked to original sources
PubMed1990

Tissue extraction of DNA and RNA and analysis by the polymerase chain reaction.

Several DNA extraction techniques were quantitatively and qualitatively compared using both fresh and paraffin wax embedded tissue and their suitability investigated for providing DNA and RNA for the polymerase chain reaction (PCR). A one hour incubation with proteinase K was the most efficient DNA extraction procedure for fresh tissue. For paraffin wax embedded tissue a five day incubation with proteinase K was required to produce good yields of DNA. Incubation with sodium dodecyl sulphate produced very poor yields, while boiling produced 20% as much DNA as long enzyme digestion. DNA extracted by these methods was suitable for the PCR amplification of a single copy gene. Proteinase K digestion also produced considerable amounts of RNA which has previously been shown to be suitable for PCR analysis. A delay before fixation had no effect on the amount of DNA obtained while fixation in Carnoy's reagent results in a much better preservation of DNA than formalin fixation, allowing greater yields to be extracted.

DNA↗
PubMed1988

[Is the renin-angiotensin system implicated in the hypertensive effect of a water-soluble tissue extract?].

A deproteinized hydrosoluble splenic extract, which produces a hypertensive effect, injected in rats intravenously is described. The pressure action is very similar to the synthetic angiotensin II, and the (Sarcosine1-Isoleucine8)--angiotensin II, a competitive antagonist, produced partial inhibition of both responses. There were no significant differences between control and experimental rats in the plasmatic levels of angiotensin II, aldosterone and ADH. Therefore, the splenic extract does not seem to release these hormones included in the renin-angiotensin pathway. Significant Angiotensin II levels were detected in the splenic material. These results support the view that forty per cent of the pressure action is due to Angiotensin II present in the extract.

Aldosterone↗
PubMed2004

A nonradioactive assay for nitric oxide synthase activity in tissue extracts.

We describe a fluorescence assay for nitric oxide synthase activity based on a new indicator, 4,5-diaminofluorescein (DAF-2). The method offers the advantage of being safer and more convenient than the citrulline radioassay in common use. The rapid and irreversible binding of DAF-2 to oxidized nitric oxide (NO) enables NO production to be measured in real time. The protocol is applied to the measurement of nitric oxide synthase in crude extracts of skeletal muscle.

Animals↗