ABSTRACT
The properties of human DNA fingerprints detected by multilocus polymeric monocore probes (PMC probes) have been investigated. The PMC probes were produced by the polymerase chain reaction with two partly complementary oligonucleotides homologous to various minisatellite or microsatellite core sequences (Ijdo J, Wells RA, Baldini A, Reeders ST. Nucleic Acids Res 1991;19:4780). It has been shown that these probes possess increased sensitivity, they detect considerably more hypervariable fragments in genomic DNA thus exhibiting advantages over the corresponding oligonucleotides and natural polycore minisatellite probes. Variation in the DNA fingerprints of different individuals produced by these probes indicates that the probability of accidental identity is very low (< 10(-12)). According to the data of cross-hybridization with PMC probes of various specificity, several distinct families can be distinguished among G-rich hypervariable sequences of the human genome.
Subject(s)
DNA Fingerprinting/methods , Oligonucleotide Probes , Bacteriophage M13/metabolism , Humans , Sensitivity and SpecificityABSTRACT
An oligonucleotide (m5C-am2A-m5C)5 containing 2'-amino-deoxyadenosine (am2A) and 5-methyldeoxycytidine (m5C) residues has been synthesized and compared with unsubstituted pentadecadeoxyribonucleotide (CAC)5 as a hybridization probe for DNA fingerprinting. It was shown that considerably higher sensitivity can be achieved with the modified analog.
Subject(s)
2-Aminopurine/analogs & derivatives , Cytosine/analogs & derivatives , DNA Fingerprinting/methods , DNA/genetics , Oligonucleotide Probes , 5-Methylcytosine , Adenine , Base Sequence , Blotting, Southern/methods , DNA/blood , DNA/isolation & purification , DNA Primers , Humans , Molecular Sequence Data , Sensitivity and SpecificityABSTRACT
The DNA sequence (AT)26 of barley genome has been cloned. This sequence is arranged in intraspecific locus and is repeated 1500 times per haploid genome. This fragment is not translated and can form cruciform structures in the AT region.
Subject(s)
DNA/genetics , Hordeum/genetics , Repetitive Sequences, Nucleic Acid , Base Sequence , Cloning, Molecular , DNA/metabolism , DNA, Superhelical/metabolism , Deoxyribonucleases, Type II Site-Specific/metabolism , Endodeoxyribonucleases/metabolism , Molecular Sequence Data , Nucleic Acid Hybridization , Polymorphism, Restriction Fragment Length , Single-Strand Specific DNA and RNA Endonucleases/metabolismABSTRACT
Hypervariable polymorphic patterns were detected with M13 phage DNA as a probe in genomic DNA of organisms belonging to different taxonomic groups including animals (vertebrates and invertebrates), plants and microorganisms. Individual-specific restriction pattern analysis (DNA fingerprinting) with this probe proved to be useful for individual identification, analysis of somatic stability and paternity testing in man. The nuclear type of inheritance indicates that the hypervariable DNA regions in question are located in the chromosomes, not in the mitochondrial DNA. The data obtained also demonstrate a potential range of M13 DNA applications as a probe for DNA fingerprinting of animals, plants and microorganisms, particularly for the determination of inbred lines, identification of bacterial strains and establishing stock, variety and strain distinctions.
Subject(s)
DNA, Viral/genetics , DNA/genetics , Polymorphism, Genetic , Animals , Coliphages/genetics , DNA/isolation & purification , DNA Restriction Enzymes , Escherichia coli/genetics , Female , Humans , Male , Paternity , Pedigree , Plants/genetics , Species SpecificityABSTRACT
Partially purified human procollagen mRNAs have been copied using reverse transcriptase from avian myeloblastosis virus. Under standard conditions, using a high deoxynucleotide concentration, only incomplete cDNAs corresponding to one-fifth to one-half of the template could be synthesized. Addition of a ribonuclease inhibitor from human placenta in the reaction mixture allowed the synthesis of full-length cDNA copies from extremely long procollagen mRNA.
