In-gel hybridization of DNA separated by pulsed field agarose gel electrophoresis
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چکیده
منابع مشابه
Pulsed-field gel electrophoresis of circular DNA.
Mobility of supercoiled (form I) and nicked circular (form II) plasmid DNAs was determined on two major forms of pulsed-field electrophoresis, CHEF and OFAGE. Plasmids with molecular lengths ranging from 2.30 to 17.8 kilobase pairs (kb) were used with Saccharomyces cerevisiae chromosomes as standards. Agarose gel concentrations were varied from 0.3 to 2.0 percent, with higher percentage gels re...
متن کاملDetermination of triglycerides in lipoproteins separated by agarose gel electrophoresis.
We developed a simple method for the quantitation of triglycerides in electrophoretically separated lipoproteins by specific enzymatic staining. After electrophoresis, glycerol is liberated from triglycerides by the action of cholesterol esterase. Glycerol is oxidized by a sequence of enzymatic reactions. Due to the presence of triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogen...
متن کاملPulsed-field gel electrophoresis.
DNA molecules in an agarose matrix elongate and migrate toward the anode when exposed to an electric field. An agarose matrix represents a highly irregular network with pores of various dimensions, large open areas, and regions with different densities. During electrophoresis, migration will be retarded by obstructions that prevent sieving by size and result in "end-on" migration as if through ...
متن کاملQuantitative hybridization to genomic DNA fractionated by pulsed-field gel electrophoresis.
Hybridization to genomic DNA fractionated by CHEF electrophoresis can vary >100-fold if the DNA is acid depurinated prior to Southern blotting. The level of hybridization is high or low depending on whether the molecule being analyzed migrates at a size coincident with or different from the size of the majority of genomic DNA in the sample, respectively. Techniques that avoid acid depurination ...
متن کاملAgarose gel electrophoresis.
MATERIALS 6x Gel-loading buffer Agarose solutions (please see Step 3) DNA samples DNA size standards Samples of DNAs of known size are typically generated by restriction enzyme digestion of a plasmid or bacteriophage DNA of known sequence. Alternatively, they are produced by ligating a monomer DNA fragment of known size into a ladder of polymeric forms. DNA staining solution For a discussion of...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1990
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/18.10.3098