Quantitative 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 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.
Leach
T J
TJ
Laboratory of Developmental Genetics, Wadsworth Center, New York State Department of Health, Albany, NY, USA.
Glaser
R L
RL
D000143:Acids / Q000494:pharmacology; D000818:Animals; D015139:Blotting, Southern / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D004262:DNA Restriction Enzymes; D004276:DNA, Satellite / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020161:Physical Chromosome Mapping / Q000379:methods; D011687:Purines / Q000378:metabolism; D014466:Ultraviolet Rays
D000143:Acids; D004276:DNA, Satellite; D011687:Purines; D004247:DNA; D004262:DNA Restriction Enzymes