A cross-hybridization method for DNA mapping with photobiotin-labeled probes.
An efficient procedure combining the Southern Cross method described by H. Potter and D. Dressler (1986, Gene 48, 229-239) and the use of nonradioactive reporter molecules which allows fast restriction mapping of DNA molecules up to 40 kb is described. In this method, photobiotin-labeled fragments for one restriction enzyme cross-hybridize with at least five unlabeled digests transferred onto nylon membranes. Hybridization spots are revealed on recipient membranes at a homologous fragment cross, allowing direct restriction map construction. In comparison with 32P "cross-blot" hybridization, photobiotin labeling is safer, faster, and easier to dispose of.
Chetrit
P
P
Laboratoire de Génétique Moléculaire des Plantes, Université de Paris-Sud, Orsay, France.
Gaudin
V
V
de Courcel
A
A
Vedel
F
F
D000345:Affinity Labels; D001386:Azides; D001710:Biotin / Q000031:analogs & derivatives*; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004587:Electrophoresis, Agar Gel; D009693:Nucleic Acid Hybridization
D000345:Affinity Labels; D001386:Azides; D015342:DNA Probes; D001710:Biotin; D004247:DNA; C045549:photobiotin