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At least 397 records · Page 22Linked to original sources
PubMed1998

[Electron microscopic in situ hybridization of dUTP-digoxigenin labeled probes with polytene chromosomes of Drosophila melanogaster].

Using DNA probes labeled with digoxygenin-11-dUTP, a simplified method of electron microscopic (EM) in situ hybridization was developed for standard squashes of Drosophila melanogaster polytene chromosomes. The developed method is efficient and reproducible: its high resolution and specificity was shown for the transformed strain 148, in which the insertion was localized by EM as a new thin band. The method was applied for fine mapping of the developmentally regulated complex gene, muscleblind (mbl), which was shown to cover the 54B1-2 large band and the adjacent interbands in 2R polytene chromosome.

Animals↗
PubMed2005

RT-PCR-based dot blot hybridization for the detection and differentiation between porcine epidemic diarrhea virus and transmissible gastroenteritis virus in fecal samples using a non-radioactive digoxigenin cDNA probe.

Multiplex reverse transcription-polymerase chain reaction (RT-PCR)-based dot blot hybridization was developed to increase the sensitivity for the detection and differentiation between porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) in fecal samples. Fecal samples found positive by RT-PCR-based agarose gel electrophoresis were always found positive by RT-PCR-based dot blot hybridization. In addition, 5 out of 10 fecal samples which were negative for PEDV by RT-PCR-based agarose gel electrophoresis were positive for PEDV by RT-PCR-based dot blot hybridization. This RT-PCR-based dot blot hybridization increased 1000-fold in sensitivity for PEDV and 100-fold for TGEV; weakly positive bands in the agarose gel electrophoresis gave a clear positive result with dot blot hybridization.

Animals↗
PubMed2001

Enzyme-amplified aequorin-based bioluminometric hybridization assays.

The sensitivity of aequorin-based bioluminometric hybridization assays was enhanced by introducing, enzymically, multiple aequorin labels per DNA hybrid. The target DNA was hybridized in microtiter wells with an immobilized capture probe and a digoxigenin-labeled detection probe. The hybrids were reacted with an anti-digoxigenin antibody conjugated to horseradish peroxidase. Peroxidase catalyzed the oxidation of digoxigenin-tyramine by hydrogen peroxide, resulting in the attachment of multiple digoxigenin moieties to the solid phase. Aequorin-labeled anti-digoxigenin antibody was then allowed to bind to the immobilized digoxigenins. The bound aequorin was determined by its characteristic Ca2+-triggered bioluminescence. As low as 20 fmol/L (1 amol/ well) target DNA was detected with a signal-to-background ratio of 2.7. A hybridization assay that used only aequorin-labeled anti-digoxigenin antibody without the peroxidase amplification step gave a signal-to-background ratio of 2 for 160 fmol/L target DNA. The signal enhancement of the amplified assay was in the range of 14-38 times. The analytical range of the amplified assay extended up to 2600 fmol/L. The CVs were in the range of 5.5-7.3%.

Aequorin↗
PubMed1980

Digoxin degradation in acidic dissolution medium.

The release of digoxin and its simultaneous conversion to digoxigenin bisdigitoxoside, digoxigenin monodigitoxoside, and digoxigenin in a USP dissolution test medium were followed by high-pressure liquid chromatography. Two products, Tablets A and B, were manufactured by solvent deposition and simple blending methods, respectively. Tablet A released digoxin faster than Tablet B in distilled water and in artificial intestinal juice, and no decomposition was observed. In the USP dissolution test medium, the rate of hydrolysis to digoxigenin bisdigitoxoside was almost equal to that of hydrolysis to digoxigenin monodigitoxoside, and a comparatively large formation rate of digoxigenin was observed. Concentrations of digoxin and its decomposition products were described by differential equations that included dissolution rates of digoxin (rapidly dissolving digoxin and digoxin crystals) and an apparent hydrolysis rate. In the earlier stage of dissolution, hydrolysis was rate determining; in the later stage, dissolution became the rate-determining step for overall digoxin degradation. To suppress digoxin hydrolysis in the USP dissolution test medium, a developmental formulation study was performed. The incorporation of magnesium oxide and magnesium hydroxide-aluminum hydroxide in the tablet formulations inhibited digoxin hydrolysis by 15.3 and 14.5%, respectively, after dissolution for 30 min without serious delay of drug release.

Acids↗