Hybridization and Detection of Biotinylated Oligonucleotide Probes in Agarose Gels
نویسندگان
چکیده
منابع مشابه
Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.
We describe a technique for transferring electrophoretically separated bands of RNA from an agarose gel to paper strips. The RNA is coupled covalently to diazobenzyloxymethyl groups on the paper. After transfer and appropriate treatment of the paper to destroy remaining diazo groups, specific RNA bands can be detected by hybridization with 32P-labeled DNA probes followed by autoradiography. Thi...
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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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Nucleic acids that have been subjected to electrophoresis through agarose gels may be detected by staining and visualized by illumination with 300-nm UV light. Methods for staining and visualization of DNA using either ethidium bromide or SYBR Gold are described here. The most convenient and commonly used method to visualize DNA in agarose gels is staining with the fluorescent dye ethidium brom...
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Specific DNA restriction endonuclease fragments can be identified after electrophoresis in agarose gels by hybridization in the gel (in situ) to radioactive homologous RNA. RNA-DNA hybrids are detected by autoradiography of the gel. Comparison of band patterns of the autoradiogram and the ethidium bromide stained gel allows the identification of the DNA fragment which is complementary to the RN...
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A dot blot hybridization test was developed for the detection of enterotoxigenic E. coli without the use of radioisotopes. Three biotin-labeled DNA (Bio-DNA) probes corresponding to structural genes specifying heat-labile and heat-stable enterotoxins of porcine and human origin were prepared by random priming; label incorporation was significantly higher than that obtained from the use of nick ...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1997
ISSN: 0736-6205,1940-9818
DOI: 10.2144/97234bm17