Denhardt solution

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Denhardt's solution is a solution used in biochemistry .

properties

The Denhardt solution is used in a Southern blot , a Northern blot , a DNA microarray or a fluorescence in situ hybridization to block unspecific binding sites for nucleic acids on the blot membrane or the microarray or the tissue section . It is added to both blocking and hybridization solutions. To increase the viscosity , it contains Ficoll (type 400, 0.1–0.2 g / L) as well as 0.1–0.2 g / L polyvinylpyrrolidone and 0.1–0.2 g / L bovine serum albumin as blocking reagents . Due to the protein content of the albumin, the solution, which is made up as a concentrated stock solution, is sterile-filtered . With a Southern blot, heat-denatured DNA from salmon or herring sperm is added to the buffer , which contains the surfactant sodium lauryl sulfate , in addition to the Denhardt solution .

Individual evidence

  1. ^ DT Denhardt: A membrane-filter technique for the detection of complementary DNA. In: Biochemical and biophysical research communications. Volume 23, Number 5, June 1966, pp. 641-646, PMID 5963888 .
  2. ^ A b Ralph Rapley: The Nucleic Acid Protocols Handbook. Springer Science & Business Media, 2000, ISBN 978-0-896-03459-4 , pp. 164, 986.
  3. ↑ Thomas Liehr: Fluorescence In Situ Hybridization (FISH) - Application Guide. Springer Science & Business Media, 2008, ISBN 978-3-540-70581-9 , p. 325.