Genomic amplification by the polymerase chain reaction (PCR) was used to identify a unique genomic sequence of Ehrlichia risticii directly in DNA isolated from peripheral-blood buffy coat cells of E. risticii-infected horses (Potomac horse fever) and from infected cell cultures. A specific primer pair, selected from a cloned, species-specific, 1-kb DNA fragment of the E. risticii genome as a te...

background pcr is a molecular technique for herpes simplex virus (hsv) detection that can cause life-threatening infections such as encephalitis and keratitis. however, the main issues, false-negative results causing by pcr inhibitors, of this technique that reduce pcr efficiency. to overcome this problem, a competitive internal amplification control (iac) was constructed for conventional pcr u...

BACKGROUND PCR is a molecular technique for herpes simplex virus (HSV) detection that can cause life-threatening infections such as encephalitis and keratitis. However, the main issues, false-negative results causing by PCR inhibitors, of this technique that reduce PCR efficiency. To overcome this problem, a competitive internal amplification control (IAC) was constructed for conventional PCR u...

The full potential of diagnostic PCR is limited, in part, by the presence of inhibitors in complex biological samples that reduce the amplification efficiency. Therefore, different pre-PCR treatments are being used to reduce the effects of PCR inhibitors. The aim of the present study was to investigate the effects of 16 amplification facilitators to enhance DNA amplification in the presence of ...

A direct, non-radioactive method of quantitative PCR amplification has been investigated for the diagnosis of deletion and duplication carriers in the dystrophin gene. The simultaneous amplification of two loci, or several loci using multiplex PCR, allows for the direct comparison of relative amounts of products from normal homozygous loci and potentially heterozygous deleted/duplicated loci. S...

The real-time polymerase chain reaction (PCR) uses fluorescent reporter molecules to monitor the production of amplification products during each cycle of the PCR reaction. This combines the DNA amplification and detection steps into one homogeneous assay and obviates the need for gel electrophoresis to detect amplification products. Appropriate data analysis and/or use of apposite chemistries ...

DNA amplification by polymerase chain reaction (PCR) should be inhibited if the target for amplification region in the template DNA is nicked or cut. Based on this premise, we established a sensitive and differential assay using PCR to detect antibiotics that act on DNA. After template lambda DNA (10 pg) was incubated with antibiotics (10 approximately 20 ng) at 37 degrees C for 30 minutes in a...

aptamers, or single stranded oligonucleotides, are produced by systematic evolution of ligands by exponential enrichment, abbreviated as selex. in the amplification and regeneration step of selex technique, dsdna is conversed to ssdna. asymmetric pcr is one of the methods used for the generation of ssdna. the purpose of this study was to design a random dna library for selection of aptamers wit...

PCR is widely employed as the initial DNA amplification step for genetic testing and cancer biomarker detection. However, a key limitation of PCR-based methods, including real-time PCR, is the inability to selectively amplify low levels of variant alleles in a wild-type allele background. As a result, downstream assays are limited in their ability to identify subtle genetic changes that can hav...

Transposons have been used in many laboratories worldwide as a powerful tool for insertional mutagenesis to investigate gene function in bacteria (8). The technique has recently been adapted in which a transposon, such as mini-Tn5, delivers a unique identifying sequence of 40 nucleotides (1). This technique, termed signature-tagged mutagenesis (STM), is widely used in the search for virulence-a...