The production of informative random amplified polymorphic DNA (RAPD) markers using PCR and a single primer is often accompanied by the generation of artifactual (noninformative) bands as well. When RAPD data are used to compute genetic similarity coefficients, these artifacts (false positives, false negatives, or both) can cause large biases in the numerical values of the coefficients. As a re...

We have isolated cDNA clones for the large etc chain of the rat brain Clathrin Associated Protein complex AP-2. A rat brain XZAP-I1 cDNA library (Stratagene) was screened with the synthetic oligonucleotide probe 5'-C(C, T) TCCAT (A ,G) TGNCC (A, G) AA (A, G) TC (G, T) ATGTCGTG, corresponding to the stretch of amino acids between His-70 and AIa-79 that are shared by both mouse aa and ac chains (...

Despite the large number of genes that are expected to be involved in non-syndromal, recessive deafness, only a few have been cloned. One of these genes is GJB2, which encodes connexin 26. A frameshift mutation in this gene has been reported to be common in several populations and a carrier frequency of about 1 in 30 people has been detected in Italy and Spain. Mutation 35delG is difficult to d...

Anthrax is a fatal infection of humans and livestock that is caused by the gram-positive bacterium Bacillus anthracis. The virulent strains of B. anthracis are encapsulated and toxigenic. In this paper we describe the development of a PCR technique for identifying spores of B. anthracis. Two 20-mer oligonucleotide primers specific for the capB region of 60-MDa plasmid pXO2 were used for amplifi...

Two recombinant DNA clones, pMG286.2 and pMG301.1, were isolated from the partial genomic library of Mycoplasma gallisepticum strain S6. Recombinant M. gallisepticum specific fragments were used as probes in Southern hybridisation with 10 M. gallisepticum strains whose DNA was digested by EcoRI, HindIII, BglII, RsaI and BamHI. The 1.5 kb fragment pMG301.1 did not show polymorphism in hybridisat...

Mycoplasma hyopneumoniae is the primary agent of mycoplasmal pneumonia in swine. In this study we performed an arbitrarily primed PCR (AP-PCR) analysis, in which low-stringency amplification with a single primer was used, to investigate genetic variability in M. hyopneumoniae strains and field isolates. We performed preliminary experiments to examine the efficacy of 40 different 10-mer oligonuc...

Division of Experimental Therapy, The Netherlands Cancer Institute Antonin van Leeuwenhoek Huis, 1066 CX Amsterdam, The Netherlands The search for sequence variations in genomic DNA becomes increasingly important in the study of inherited disease genes as well as of genes that play a role in the development of cancer. A wide variety of different methods to detect DNA sequence variations has bee...

Eukaryotic initiation factor 5A (eIF-5A) is the only cellular protein known to contain a hypusine residue that is formed by transferring the aminobutyl moiety from spermidine to a specific lysine residue, followed by hydroxylation at the aminobutyl group. A simple PCR-based strategy was developed to obtain a full-length cDNA of Neurospora crassa eIF-5A. The strategy consists of (i) the design o...

The polymerase chain reaction was used to obtain randomly amplified polymorphic DNA profiles for genetic fingerprinting of twelve different isolates of Staphylococcus aureus from Riyadh region, KSA using RAPD-PCR technique eleven out of forty operon primers (10-mer primers) showed polymorphism among the isolates tested generating over 65 bands, approximately 42 of which were polymorphic with si...

Erwinia amylovora, the causative agent of fire blight, was identified independently from the common plasmid pEA29 by three different PCR assays with chromosomal DNA. PCR with two primers was performed with isolated DNA and with whole cells, which were directly added to the assay mixture. The oligonucleotide primers were derived from the ams region, and the PCR product comprised the amsB gene, w...