Journal Article

Quantitative hybridization to genomic DNA fractionated by pulsed-field gel electrophoresis

Thomas J. Leach and Robert L. Glaser

in Nucleic Acids Research

Volume 26, issue 20, pages 4787-4789
Published in print October 1998 | ISSN: 0305-1048
Published online October 1998 | e-ISSN: 1362-4962 | DOI: https://dx.doi.org/10.1093/nar/26.20.4787
Quantitative hybridization to genomic DNA fractionated by pulsed-field gel electrophoresis

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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 including in-gel hybridizations and UV irradiation of DNA prior to blotting provide more accurate quantitative results. CHEF analysis of DNA molecules containing repetitive satellite sequences is particularly prone to this effect.

Journal Article.  2025 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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