Optimized oligonucleotide probes for DNA fingerprinting

Schäfer, Renate; Zischler, Hans; Birsner, U.; Becker, Andrea C.; Epplen, Jörg T.

doi:10.1002/elps.1150090804

Cited by 145 publications

(34 citation statements)

References 29 publications

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“…Genomic DNA was isolated from brain, liver, spleen and gill according to Blin and Stafford (1976). DNA (5 Jlg) from all individuals was digested with restriction endonucleases (Haeiii, Hinfi, Alu!, Sau3AI) according to the recommendations of the supplier (Boehringer· Mannheim, FRG) and resolved on 0.8% horizontal agarase gels in TAE buffer (40 mM Tris, 12 mM sodium acetate, 2 mM EDTA, pH 8.3) at 2 V/ern for about 40 h. Prior to hybridization the gels were dry-blotted, stained with ethidium bromide, photographed, denatured, neutralized and reequilibrated as described (Schäfer et al 1988). The oligonucleotide probes specific for simple repeats were synthesized on an automated DNA synthesizer (Applied Biosystems 381A, Weiterstadt, FRG) and were end-labelled with [y 32 P)ATP (Amersham, Braunschweig, FRG) in a standard kinase reaction.…”

Section: Methodsmentioning

confidence: 99%

Early stages of sex chromosome differentiation in fish as analysed by simple repetitive DNA sequences

Nanda

Schartl²,

Feichtinger

et al. 1992

Chromosoma

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Abstract. Animal sex chromosome evolution has started on different occasions with a homologous pair of autosomes leading to morphologically differentiated gonosomes. In contrast to other vertebrate classes, among fishes cytologically dernonstrahle sex chromosomes are rare. In reptiles, certain motifs of simple tandemly repeated DNA sequences like (gata)n/(gaca)m are associated with the constitutive heterochromatin of sex chromosomes. In this study a panel of simple repetitive sequence probes was hybridized to restriction enzyme digested genomic DNA of poeciliid fishes. Apparent male heterogamety previously established by genetic experiments in Poecilia reticulata (guppy) was correlated with male-specific hybridization using the (GACA) 4 probe. The (GAT A) 4 oligonucleotide identifies certain male guppies by a Y chromosomal polymorphism in the outbred population. In cantrast none of the genetically defined heterogametic situations in Xiphophorus could be verified consistently using the collection of simple repetitive sequence probes. Only individuals from particular populations produced sex-specific patterns of hybridization with (GAT A) 4 • Additional poeciliid species (P. sphenops, P. velifera) harbour different sex-specifically organized simple repeat motifs. The observed sex-specific hybridization patterns were substantiated by banding analyses of the karyotypes and by in situ hybridization using the (GACA) 4 probe.

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Section: Methodsmentioning

confidence: 99%

Early stages of sex chromosome differentiation in fish as analysed by simple repetitive DNA sequences

Nanda

Schartl²,

Feichtinger

et al. 1992

Chromosoma

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“…I n this experiment, a n oligonucleotide .probe gp 2000 (CTCCACCT)3, prepared by authors (Fang et al, 2002), was 5'endlabelled with [3zP] using T4 polynucleotide kinase according to the protocol of Schaer et al (1988).…”

Section: S O II T H Erii B 1 O T 1 I I Igmentioning

confidence: 99%

Genetic Diversity of the Giant Panda(Ailuropoda melanoleuca)Between Big and Small Populations

Fang

Wan

Fujihara

2002

Journal of Applied Animal Research

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“…'' sex differences depending on tbe population investigated "'associated with nucleolus organizers 57 genetic individualization in the human was subsequently achieved in a single hybridization step using (CAC) 5 (Schäfer et al, 1988). Solely depending on the resolving power of the gel system used, all human individuals can be distinguished with the only exception of genetically identical, monozygotic twins.…”

Section: Oligonucleotide Fingerprinting In Man With Special Referencementioning

confidence: 99%

Oligonucleotide Fingerprinting Using Simple Repeat Motifs: A Convenient, Ubiquitously Applicable Method to Detect Hypervariability for Multiple Purposes

Epplen¹,

Ammer²,

Epplen³

et al. 1991

Experientia Supplementum

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A panel of simple repetitive oligonucleotide probes has been designed and tested for multilocus DNA fingerprinting in some 200 fungal, plant and animal species as well as man. To date at least one of the probes has been found to be informative in each species. The human genome, however, has been the major target of many fingerprintins studies. Using the probe (CAC) 5 or (GTG) 5 , individualization of all humans is possible except for monozygotic twins. Paternity analyses are now perfonned on a routine basis by the use of multilocus fingerprints, inctuding also cases of deficiency, i.e. where one of the parents is not available for analysis. In forensie science stain analysis is feasible in all tissue remains containing nuc)eated cells. Depending on the degree of DNA degradation a variety of oligonucleotides are informative, and they have been proven useful in actual case work. Advantages in comparison to other methods including enzymatic DNA amplification techniques (PCR) are evident. Fingerprint patterns of tumors may be changed due to the gain or loss of chromosomes and/or intrachromosomal deletion and amplification events. Locus-specific probes were isolated from the human (CAC) 5 /(GTG) 5 fingerprint with a varying degree of informativeness (monomorphic versus truly hypervariable markers). The feasibility of three different approaches. for the isolation of hypervariable mono-locus probes was evaluated. Finally, one particular mixed simple (gt)n(ga)m repeat locus in the second intron of the HLA-DRB genes has been scrutinized to allow comparison of the extent of exon-encoded (protein-) polymorphisms versus intronie bypervariability of simple repeats: adjacent to a single gene sequence (e.g. HLAMDRBl•0401) many different length alleles were found. Group-specific structures of basic repeats were identified within the evolutionarily related DRB alleles. As a further application it is suggested here that due to the ubiquitous interspersion of their targets, short probes for simple repeat sequences are especially useful tools for ordering genomic cosmid, yeast artificial chromosome and phage banks.

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Optimized oligonucleotide probes for DNA fingerprinting

Cited by 145 publications

References 29 publications

Early stages of sex chromosome differentiation in fish as analysed by simple repetitive DNA sequences

Early stages of sex chromosome differentiation in fish as analysed by simple repetitive DNA sequences

Genetic Diversity of the Giant Panda(Ailuropoda melanoleuca)Between Big and Small Populations

Oligonucleotide Fingerprinting Using Simple Repeat Motifs: A Convenient, Ubiquitously Applicable Method to Detect Hypervariability for Multiple Purposes

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