Plant DNA fingerprinting with radioactive and digoxigenated oligonucleotide probes complementary to simple repetitive DNA sequences.
The existence of hypervariable DNA sequences in nuclear genomes, and the use of appropriate "fingerprinting" probes to detect them, has gained widespread scientific interest, and also led to multiple applications in diverse areas. Two years ago, the new technique of "DNA fingerprinting" was also introduced into the analysis and characterization of plant genomes, initially by using human or M13 minisatellites as probes. In the present article, we demonstrate the applicability for plant DNA fingerprinting of oligonucleotide probes specific for simple repetitive DNA sequences. We show that various levels of intra- and interspecific polymorphisms can be detected; the information to be gained depends on the optimal combination of probe and species. Variety-specific patterns were obtained in several cases. Some probes revealed variability between individuals. Somatic variability was not observed. Different DNA isolation and purification procedures were tested in order to introduce a fast and easy-to-perform isolation method suitable for a large variety of plant species. Nonradioactive fingerprinting was performed using digoxigenated oligonucleotides as probes. Banding patterns obtained with radioactive and digoxigenin-based labeling techniques proved to be of similar quality.
Weising
K
K
Pflanzliche Molekularbiologie, Fachbereich Biologie, Johann Wolfgang Goethe-Universität, Frankfurt, Germany.
Beyermann
B
B
Ramser
J
J
Kahl
G
G
D001483:Base Sequence; D016172:DNA Fingerprinting; D004076:Digoxigenin; D015345:Oligonucleotide Probes; D010759:Phosphorus Isotopes; D010944:Plants / Q000235:genetics*; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid
D015345:Oligonucleotide Probes; D010759:Phosphorus Isotopes; D004076:Digoxigenin