Isolation of human minisatellite loci detected by synthetic tandem repeat probes: direct comparison with cloned DNA fingerprinting probes.
As a direct comparison with cloned 'DNA fingerprinting' probes, we present the results of screening an ordered array Charomid library for hypervariable human loci using synthetic tandem repeat (STR) probes. By recording the coordinates of positive hybridization signals, the subset of clones within the library detected by each STR probe can be defined, and directly compared with the set of clones detected by naturally occurring (cloned) DNA fingerprinting probes. The STR probes vary in the efficiency of detection of polymorphic minisatellite loci; among the more efficient probes, there is a strong overlap with the sets of clones detected by the DNA fingerprinting probes. Four new polymorphic loci were detected by one or more of the STR probes but not by any of the naturally occurring repeats. Sequence comparisons with the probe(s) used to detect the locus suggest that a relatively poor match, for example 10 out of 14 bases in a limited region of each repeat, is sufficient for the positive detection of tandem repeats in a clone in this type of library screening by hybridization. These results not only provide a detailed evaluation of the usefulness of STR probes in the isolation of highly variable loci, but also suggest strategies for the use of these multi-locus probes in screening libraries for clones from hypervariable loci.
Armour
J A
JA
Department of Genetics, University of Leicester, UK.
Vergnaud
G
G
Crosier
M
M
Jeffreys
A J
AJ
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D016172:DNA Fingerprinting; D015342:DNA Probes; D004276:DNA, Satellite / Q000235:genetics*; D005069:Evaluation Studies as Topic; D015723:Gene Library; D006801:Humans; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid; D012689:Sequence Homology, Nucleic Acid
D015342:DNA Probes; D004276:DNA, Satellite; D004247:DNA