Quantitative in situ hybridization with enhanced sensitivity in soft, bone and tooth tissue using digoxigenin tagged RNA probes
نویسندگان
چکیده
منابع مشابه
Hybridization and detection of digoxigenin probes on RNA blots.
This article describes nonradioactive probing of a Northern blot. The method employs digoxigenin-labeled probes. Antidigoxigenin antibody/alkaline phosphate conjugate, and a chemiluminescent substrate are subsequently used in the detection system.
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Detection of transcripts in situ is a rapid means by which gene expression can be characterized in many systems. In the nematode, Caenorhabditis elegans, the ease with which transgenics can be made and the general reliability of reporter fusion expression patterns, have made this technique comparatively less popular than in other systems. There are, however, still applications in which in situ ...
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The sensitivity of digoxigenin and biotin labelled DNA probes for the detection of human papillomavirus (HPV) by dot blotting and in situ hybridisation was compared in tissues from cervical, laryngeal, and anogenital neoplasia. Probes were either labelled with digoxigenin by the random primer technique and detected with anti-digoxigenin antibody, or labelled with biotin by nick translation and ...
متن کاملA Small-scale Northern Hybridization with Digoxigenin- labeled RNA Probes from PCR-based Templates
CONTENTS Abstract We describe the use of PCR to generate sense and antisense template DNA for nonradioactive in vitro transcription reactions. The resulting digoxigenin-labeled RNA probes can be used in a small-scale Northern hybridization procedure that is performed in microcentrifuge tubes. This kind of Northern approach is especially useful when the expression of several genes is to be initi...
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Equipment and reagents • Glass coverslips, 22x22mm • 6-well plates • 4% freshly made paraformaldehyde (Electron Microscopy Sciences) in PBS, pH 7.4 • Triton X-100 (Sigma) • Parafilm • Forceps, Dumont, GG (Electron Microscopy Sciences) • Avidin-DCS-Texas Red or –fluorescein (Vector) • Filter paper • Mounting medium (Molecular Probes) • Microscopy coverslide • Deionized formamide (Ambion) • Yeast...
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ژورنال
عنوان ژورنال: Biochemia Medica
سال: 2008
ISSN: 1846-7482
DOI: 10.11613/bm.2008.008