Source and Description of Primers. The initial primers for the PCR were designed based on human DNA sequence (accession no. X92412; Kolmerer et al., 1996). The position of the forward and reverse primers corresponded to exon 3 and exon 5, respectively. These primers are expected to amplify a fragment of 1.93 kb from human DNA. A PCR fragment of 2.1 kb was amplified from porcine genomic DNA. Par...

In conventional DNA fingerprinting, hypervariable and repetitive sequences (minisatellite or microsatellite DNA) are detected with hybridization probes. As demonstrated here, these probes can be used as single primers in the polymerase chain reaction (PCR) to generate individual fingerprints. Several conventional DNA fingerprinting probes were used to prime the PCR, yielding distinctive, hyperv...

In vitro DNA amplification methods, such as polymerase chain reaction (PCR), rely on synthetic oligonucleotide primers for initiation of the reaction. In vivo, primers are synthesized on-template by DNA primase. The bacteriophage T7 gene 4 protein (gp4) has both primase and helicase activities. In this study, we report the development of a primase-based Whole Genome Amplification (pWGA) method,...

Random amplified polymorphic DNA (RAPD) is a simple, fast and cost effective method for assessing genetic diversity of plant varieties. The reproducibility of the RAPD amplification was affected by several factors, therefore causing misinterpretation. In this study, we analyzed the effect of changing the concentration of magnesium chloride, template genomic DNA and Taq DNA polymerase with the o...

USER cloning is a fast and versatile method for engineering of plasmid DNA. We have developed a user friendly Web server tool that automates the design of optimal PCR primers for several distinct USER cloning-based applications. Our Web server, named AMUSER (Automated DNA Modifications with USER cloning), facilitates DNA assembly and introduction of virtually any type of site-directed mutagenes...

DNA polymerases amplify DNA fragments through primer extension reactions. However, polymerization behavior of short primers in the primer extension process has not been systematically explored. In this study, we examined the minimal primer length required for primer extension, and the effect of primer length, mismatches and other conditions on DNA polymerization using a non-radioactive method. ...

A sensitive and highly reproducible multiplexed primer extension assay is described for quantitative mutation analysis of heterogeneous DNA populations. Wild-type and mutant target DNA are simultaneously probed in competitive primer extension reactions using fluorophor-labeled primers and high fidelity, thermostable DNA polymerases in the presence of defined mixtures of deoxy- and dideoxynucleo...

Genetic variation and inheritance of randomly amplified polymorphic DNA (RAPD) markers in female grass carp (Ctenopharyngodon idella) and male bighead carp (Hypophthalmichthys nobilis) and their F1 offspring hybrid have been studied. For this purpose, genomic DNA was extracted from muscle tissue according to phenol-chloroform method, and six decamer primers were used for amplifying polymorphic ...

A nested polymerase chain reaction (PCR) was developed to detect genomic DNA of Piscirickettsia salmonis, the causative agent of an epizootic disease in salmonids. The nested PCR assay, which used general bacterial 16s rDNA primers in the flrst amplification reaction, and P salmonisspecif~c primers in a second reaction, allowed detection of less than 1 P salmonis tissue culture infect~ous dose ...

Invasive aspergillosis (1 is a life-threatening condition in immunocompromised patients. An early diagnosis is of great importance because early treatment may resolve this potentially fatal infection. Recently, the polymerase chain reaction (PCR) has been used successfully in detecting specific DNA of several pathogen. In this study, nested PCR was used to detect DNA specific for A!.pergiflus s...