Cloning and Characterisation of Novel Single Locus Probes for Forensic Purposes

Abstract

From the analysis of multi-locus probes (Jeffreys et al, 1986; Fowler et al, 1988) it has previously been demonstrated that there are numerous hypervariable loci in the human genome. Many hypervariable loci have been fortuitously identified (Wyman and White, 1980; Bell et al, 1982; Higgs et al, 1981; Capon et al, 1983). A cloning strategy, used specifically to isolate hypervariable DNA was developed by Wong et al (1986, 1987) who used lambda libraries of human DNA; the insert consisting of size selected high molecular weight (5–15kB) DNA fragments. This procedure enriched the library for tandemly repeated minisatellites. Recently, Armour et al (1990) introduced a more successful and simpler method for cloning hypervariable DNA by constructing Charomid libraries (Saito and Stark, 1986). Charomids are cosmid derived vectors which do not have the same size constraints of lambda vectors, hence the cloning of rather unstable minisatellite fragments, which tend to lose repeats, is much improved.