Characterization of a panel of highly variable minisatellites cloned from human DNA

Abstract

Five of the most variable loci detected in human DNA by hybridization with DNA fingerprint probes have been cloned and characterized. Each locus consists of a tandem-repetitive minisatellite, with repeat units ranging in length from 9 to 45 base pairs depending on the locus. All of these cloned minisatellites act as locus-specific hybridization probes, and detect extremely variable Mendelian loci with heterozygosities ranging from 90 to 99%. These five hypervariable loci, together with a previously-isolated minisatellite designated p lambda g3, are dispersed over four autosomes (chromosomes 1, 5, 7 and 12). Syntenic pairs on chromosomes 1 and 7 show no detectable pair-wise linkage, and thus these hypervariable loci show no evidence of clustering within the genome and should provide valuable markers for mapping inherited disease. The locus-specific minisatellites act as very sensitive hybridization probes, and can be pooled to detect several hypervariable loci simultaneously. The applications of these probes in individual identification, paternity testing and analysis of cell chimaerism are discussed, and are illustrated by an analysis of forensic specimens from two victims who had been sexually assaulted and murdered.

XH1 – a new cervical carcinoma cell line and xenograft model of tumour invasion, 'metastasis' and regression

“…Hypervariable minisatellite DNA probing Locus-specific hypervariable minisatellite probes, MS1 and MS31 (ICI Biological Products, Cheshire) (Wong et al, 1987; were used to ensure that the XH1 cells were not contaminated with the established cell lines Caski (Patillo et al, 1977), A431 (Giard et al, 1973) and Bowes melanoma (Rifkin et al, 1974), also maintained in this laboratory.…”

Rapid Individual Identification by Minisatellite Variant Repeat (MVR)-PCR at D1S8 Locus Using “Exponential Law”

“…Here, the appearance of two variant 29 bp repeat units at each code position is originating from the DNA of parents. Two major classes of a-type and t-type differ by a single base substitution 5 and show highly diverse dispersion patterns within alleles. 4 This MVR-PCR method is highly precise in individual identification.…”

“…3 D1S8 locus on chromosome 1 contains three classes of variant 29 bp repeat units: a-type, t-type and (rarely) unamplified null or O-type repeats. 4,5 When analyzed electrophoretically using two tracks a and t for the analysis of atype and t-type units, respectively, MVR-PCR of total genomic DNA produces a profile of both alleles superimposed to generate six different types for each "rung" of the ladder; an intense band in the a track with no band in the t track (code 1, aa; called homozygote), an intense band in the t track with no band in the a track (code 2, tt; called homozygote), a faint a band with a faint t band (code 3, at; called heterozygote), a faint a band without t band (code 4, a0; called heterozygote), a faint t band without a band (code 5, t0; called heterozygote) and no band in either a or t track (code 6, 00). Code 4 and code 5 were distinguished from code 1 and code 2 by their intensities of bands, respectively.…”

Genetic evidence that placental site trophoblastic tumours can originate from a hydatidiform mole or a normal conceptus

“…We report here two cases of PSTT in which the genetic origin has been established using a Y chromosome-specific probe CY84 (Wolfe et al, 1985) and a panel of locus-specific minisatellite probes, which identify highly polymorphic restriction fragment length polymorphisms (RFLPs) of the DNA (Wong et al, 1987;Armour et al, 1990 RFLPs of the DNA, defined by the locus-specific minisatellite probe MSI (Wong et al, 1987), were examined in HinfI digested parental and tumour DNA as previously described (Wong et al, 1987;Fisher et al, 1989). The probe was subsequently stripped from the DNA and rehybridisation carried out successively with probes MS8, MS31, p4g3, MS43 and MS621 (Wong et al, 1987;Armour et al, 1990).…”

“…The probe was subsequently stripped from the DNA and rehybridisation carried out successively with probes MS8, MS31, p4g3, MS43 and MS621 (Wong et al, 1987;Armour et al, 1990). In the case JD parental and tumour DNA was reprobed with three additional locus-specific minisatellite probes MS605, MS622 and MS620 (Armour et al, 1990).…”