title: Direct in-gel hybridization of digoxigenin-labelled non-radioactive probes.
abstract: An improved, simple, cost-effective and non-radioactive procedure for in-gel hybridization is described for the detection of signal in dried agarose gels. Large and small digoxigenin-labelled DNA and oligonucleotide probes hybridized efficiently and specifically with the complementary DNA sequences in the gel. The signal-to-noise ratios for the gels dried at 55 degrees C at 1 atmospheric pressure were 3-3.5-fold higher than the gels dried at 25 degrees C under vacuum. The method shows an increased sensitivity over currently available non-radioactive methods for in-gel hybridization. A single copy of a gene insert could be detected by the use of this procedure.
authors.affiliation: National Center for Toxicological Research, Food and Drug Administration, 3900 NCTR Road, Jefferson, AR 72079, USA.
authors.forename: S A
authors.initials: SA
authors.lastname: Khan
authors.forename: M S
authors.initials: MS
authors.lastname: Nawaz
authors.forename: A A
authors.initials: AA
authors.lastname: Khan
authors.forename: C E
authors.initials: CE
authors.lastname: Cerniglia
mesh_terms: D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D004076:Digoxigenin; D004587:Electrophoresis, Agar Gel; D005782:Gels; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012685:Sepharose
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; D004279:DNA, Viral; D005782:Gels; D012685:Sepharose; D004076:Digoxigenin