title: Effect of formaldehyde on the efficiency of hybridization of DNA immobilized on nitrocellulose filters.
abstract: We report in this study that under certain conditions formaldehyde interacts with DNA and makes it more efficient for hybridization on nitrocellulose filters. Hybridization signals of formaldehyde-treated DNA are stronger (up to 10 fold) as compared with that of the heat- or alkali-denatured DNA. Various parameters of the DNA-formaldehyde reaction are optimized as follows: (a) 6 x SSC, 10% formaldehyde, 60 degrees C, 20-30 min, reaction volume 10-200 microliters or (b) 6 x SSC, 5% formaldehyde, 98 degrees C, 15 min, reaction volume 10-200 microliters. Treatment of agarose gels after electrophoresis with formaldehyde improved both the transfer of DNA and the efficiency of hybridization. The following conditions are recommended for gel treatment: denaturation in 0.3 N NaOH, 1 M NaCl followed by neutralization with 0.5 M phosphate buffer, pH 7.0, containing 10% formaldehyde at 60 degrees C for 20 min.
authors.affiliation: Department of Botany, University of Toronto, Ontario, Canada.
authors.forename: I G
authors.initials: IG
authors.lastname: Ivanov
authors.forename: M G
authors.initials: MG
authors.lastname: AbouHaidar
mesh_terms: D003101:Collodion; D004247:DNA / Q000032:analysis* / Q000235:genetics; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005557:Formaldehyde; D007202:Indicators and Reagents; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010761:Phosphorus Radioisotopes; D010957:Plasmids
chemical_list: D007202:Indicators and Reagents; D010761:Phosphorus Radioisotopes; D005557:Formaldehyde; D003101:Collodion; D004247:DNA