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New Primer Set (new + primer_set)

Distribution by Scientific Domains

Medical Sciences	50%

Selected Abstracts

An evaluation of PCR primer sets used for detection of Propionibacterium acnes in prostate tissue samples

THE PROSTATE, Issue 14 2008
Karen S. Sfanos
Abstract BACKGROUND Multiple studies have now shown that Propionibacterium acnes can be cultured from post-prostatectomy derived prostate tissue samples. In contrast, both universal eubacterial 16S rDNA PCR and P. acnes -specific 16S rDNA PCR have failed to detect this organism at a frequency similar to that of bacterial culture. A potential explanation for this discrepancy, proposed by Cohen et al., involves mismatches in 16S rDNA primer sets used for bacterial detection. METHODS The sensitivity of both a previously published P. acnes -specific primer set containing a potential mismatch and a new primer set with no mismatches was determined. Both primer sets were used to interrogate two sets of DNA samples derived from post-prostatectomy prostate tissues that differed in the level of sterile precautions maintained during tissue collection. RESULTS The number of P. acnes positive samples was associated with the sterility of the sample collection process. In all instances, positive samples were determined to reflect low cell numbers (<10 CFU). CONCLUSIONS Although the results of previous studies have shown that P. acnes is not the only organism potentially present in the prostates of prostate cancer patients, mismatches in PCR primer sets may have also influenced the sensitivity of P. acnes detection. When using PCR in determining the presence of P. acnes in the human prostate, care should be taken to establish the potential influence of exogenous contamination and, due to the sensitivity of the assay, samples exposed to the urethra during the collection process (prostatic secretions, TURP specimens) should not be used. Prostate 68: 1492,1495, 2008. © 2008 Wiley-Liss, Inc. [source]

Real-time Polymerase Chain Reaction to Follow the Response of Muscle to Training

ARTIFICIAL ORGANS, Issue 8 2008
Lauren M. Moore
Abstract:, The adaptive response of muscle to changes in activity or loading can take many weeks. Changes in the levels of RNA within a muscle fiber can give an early indication of the nature of the response of that fiber to changes in activity or loading. We have designed a new primer set for quantitative polymerase chain reaction (PCR) that will allow us to follow these early transcriptional changes in rat muscle, and have shown that analysis can be performed by standard techniques on as little as 5 mg of muscle, an amount that can be obtained by needle biopsy. [source]

Identification of potentially toxic environmental Microcystis by individual and multiple PCR amplification of specific microcystin synthetase gene regions

ENVIRONMENTAL TOXICOLOGY, Issue 3 2005
Youness Ouahid
Abstract Reliable cyanotoxin monitoring in water reservoirs is difficult because of, among other reasons, unpredictable changes in cyanobacteria biomass, toxin production, and inadequate sampling frequency. Therefore, it would be useful to identify potentially microcystin-producing strains of cyanobacterial populations in field samples. With this aim, we developed a methodology to distinguish microcystin-producing from non-producing Microcystis strains by amplifying six characteristic segments of the microcystin synthetase mcy cluster, three corresponding to the nonribosomal peptide synthetase, genes mcyA, mcyB, and mcyC, and three to the polyketide synthase, genes mcyD, mcyE, and mcyG. For this purpose five new primer sets were designed and tested using purified DNA, cultured cells, and field colonies as DNA sources. Simultaneous amplification of several genes in multipex PCR reactions was performed in this study. The results obtained showed that: (i) the expected specific amplicons were obtained with all microcystin-producing strains but not with nonproducing strains; (ii) cells could be directly used as DNA templates, 2000 cells being a sufficient number in most cases; (iii) simultaneous amplification of several gene regions is feasible both with cultured cells and with field colonies. Our data support the idea that the presence of various mcy genes in Microcystis could be used as a criterion for ascribing potential toxigenicity to field strains, and the possibility of applying whole-cell assays for the simultaneous amplification of various genes may contribute significantly to simplifying toxigenicity testing. © 2005 Wiley Periodicals, Inc. Environ Toxicol 20: 235,242, 2005. [source]

Microsatellite markers characterized in the barn owl (Tyto alba) and of high utility in other owls (Strigiformes: AVES)

MOLECULAR ECOLOGY RESOURCES, Issue 6 2009
ÁKOS KLEIN
Abstract We have identified 15 polymorphic microsatellite loci for the barn owl (Tyto alba), five from testing published owl loci and 10 from testing non-owl loci, including loci known to be of high utility in passerines and shorebirds. All 15 loci were sequenced in barn owl, and new primer sets were designed for eight loci. The 15 polymorphic loci displayed two to 26 alleles in 56,58 barn owls. When tested in 10 other owl species (n = 1,6 individuals), between four and nine loci were polymorphic per species. These loci are suitable for studies of population structure and parentage in owls. [source]