RESTRICTABLE DNA FROM SLOUGHED CETACEAN SKIN; ITS POTENTIAL FOR USE IN POPULATION ANALYSIS
ABSTRACT Several species of cetaceans naturally slough visible quantities of skin. We have investigated the prospect of using this material as a viable alternative to the use of biopsy darts for the collection of samples for molecular analysis. Pieces of skin were collected from free-ranging individuals of three different species—the humpback (Megaptera novaeangliae), sperm (Physeter macrocephalus) and right whales (Eubalaena glacialis). DNA was extracted from 11 pieces of sloughed skin and DNA “fingerprint” profiles prepared. All samples contained DNA of both sufficient quality and quantity for genetic analysis. The applicability of this approach is discussed generally in relation to answering problems about the population structure and breeding systems of cetaceans.
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ABSTRACT: Hypervariable minisatellites can be amplified from human DNA by the polymerase chain reaction, using primers from DNA flanking the minisatellite to amplify the entire block of tandem repeat units. Minisatellite alleles up to 5-10 kb long can be faithfully amplified. At least six minisatellite loci can be co-amplified from the same DNA sample and simultaneously detected to provide a reproducible and highly variable DNA fingerprint which can be obtained from nanogram quantities of human DNA. The polymerase chain reaction can also be used to analyse single target minisatellite molecules and single human cells, despite the appearance of spurious PCR products from some hypervariable loci. DNA fingerprinting at the level of one or a few cells therefore appears possible.Nucleic Acids Research 01/1989; 16(23):10953-71. · 8.28 Impact Factor
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ABSTRACT: Variation in internal minisatellite structure can be analyzed by mapping variant repeat units within amplified alleles. A system capable of distinguishing greater than 10(70) allelic states at the human hypervariable locus D1S8 has been developed. Population surveys of internal allelic structure indicate that D1S8 alleles evolve rapidly along haploid chromosome lineages. Internal mapping of deletion mutant alleles physically selected from genomic DNA provides further evidence that germline and somatic mutations altering the number of allelic repeat units seldom if ever arise by unequal exchange between alleles. The existence of low level germline mosaicism for new mutants further indicates that many germline mutation events are premeiotic. Physical selection of new mutants also allows minisatellite mutation rates to be estimated directly in human DNA.Cell 03/1990; 60(3):473-85. · 31.96 Impact Factor
- Nature 06/1988; 333(6171):305. · 38.60 Impact Factor