Direct in-gel hybridization of digoxigenin-labelled non-radioactive probes.
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.
Khan
S A
SA
National Center for Toxicological Research, Food and Drug Administration, 3900 NCTR Road, Jefferson, AR 72079, USA.
Nawaz
M S
MS
Khan
A A
AA
Cerniglia
C E
CE
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
D015342:DNA Probes; D004269:DNA, Bacterial; D004279:DNA, Viral; D005782:Gels; D012685:Sepharose; D004076:Digoxigenin