Enhanced conditions for DNA fingerprinting with biotinylated M13 bacteriophage.
Deoxyribonucleic acid (DNA) fingerprints are Southern blots which have a pattern resembling bar codes. The pattern is created by DNA probes that bind to variable-length repeated sequences of human genomic DNA digested with restriction endonucleases. To improve DNA fingerprints obtained with biotin-labeled M13mp8 replicative form (RF) bacteriophage as the gene probe, the conditions for hybridization and the subsequent washing steps of the filter were refined. Experiments were conducted varying the electrophoresis time, blotting membranes, hybridization solution, and posthybridization washes. The simplicity, sensitivity, and reliability of this nonistopic technique make possible its application for identification of individuals within a species, for parentage testing, and for monitoring bone marrow transplantation.
Moreno
R F
RF
Kansas University Medical Center, Department of Pathology and Oncology, Kansas City.
Booth
F
F
Thomas
S M
SM
Tilzer
L L
LL
D001435:Bacteriophages / Q000235:genetics*; D001710:Biotin; D015139:Blotting, Southern; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004586:Electrophoresis; D006801:Humans; D009693:Nucleic Acid Hybridization; D009710:Nucleotide Mapping / Q000379:methods*; D011237:Predictive Value of Tests; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping
D015342:DNA Probes; D001710:Biotin; D004247:DNA