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DNA for hybridization is used to block non-specific binding of probes to membranes. This product is prepared by sonication of purified herring sperm DNA. Sonication shears the large molecular weight DNA to produce fragments in a size range of 587 to 831 base pairs. This range has been shown to be the most effective for hybridizations. The material is monitored during sonication by electrophoresis in order to determine the size range. Once sonication is complete, the material is denatured by boiling. Many factors contribute to the signal-to-noise ratio in nucleic acid hybridizations. These factors include the presence of solvent (formamide), hybridization temperature, length of hybridization, volume of hybridization solution, degree and method of agitation, use of blocking reagents, concentration and specific activity of the probe, use of molecular agents to increase the rate of nucleic acid reassociation, and the degree of stringency used during the washing of the membrane. In order to decrease any non-specific hybridization of the probe to a substrate, blocking agents must be used. Generally, a combination of blocking reagent, detergent, and denatured, fragmented DNA is used to accomplish this. This product can be used as a blocking agent in Northern and Southern blotting as well as Chromatin Immunoprecipitation and other nucleic acid hybridization techniques.
Our Abpromise guarantee covers the use of ab46666 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent dilution.|
|In situ hybridization||Use at an assay dependent dilution.|
|Blocking||Use at an assay dependent dilution.|