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PCR Primer Sets (pcr + primer_set)

Distribution by Scientific Domains
Life Sciences75%

Selected Abstracts

Evidence for cyanophages active against bloom-forming freshwater cyanobacteria

FRESHWATER BIOLOGY, Issue 6 2008
LI DENG
Summary 1. A total of 35 putative cyanophages able to infect non-axenic cultures of bloom-forming freshwater cyanobacteria in the genera Microcystis, Anabaena and Planktothrix were isolated from Lake Zurich (Switzerland) and lakes in the Cotswold Water Park (U.K.). Eleven lytic cyanophage isolates were isolated on Microcystis and 12 each on Anabaena and Planktothrix. Cyanophage isolation protocols varied when using these different cyanobacterial hosts. 2. The collection of putative cyanophage isolates encompassed a variety of morphotypes, including the first filamentous cyanophage from any environment and the second siphocyanophage reported from fresh water. 3. PCR primer sets for gp20, gp23 and MCP genes, which have been previously found to be conserved in other cyanophages, were used in an attempt to determine genetic diversity among the phage isolates. The failure to obtain specific amplification products from most isolates suggests that the cyanophages isolated in this study were different from those previously characterized from both marine and freshwater environments. 4. Some putative cyanophages within the collection of isolates proved to have a very broad host range and were able to infect Anabaena, Microcystis and Planktothrix. The ability to infect a wide range of host taxa extends the potential reproductive period for lytic propagation, and also has implications for the transfer of genetic information between deeply separated cyanobacterial lineages. [source]

Evaluation of the PCR method for identification of Bifidobacterium species

LETTERS IN APPLIED MICROBIOLOGY, Issue 1 2008
S.Y. Youn
Abstract Aims:,Bifidobacterium species are known for their beneficial effects on health and their wide use as probiotics. Although various polymerase chain reaction (PCR) methods for the identification of Bifidobacterium species have been published, the reliability of these methods remains open to question. Methods and Results:, In this study, we evaluated 37 previously reported PCR primer sets designed to amplify 16S rDNA, 23S rDNA, intergenic spacer regions, or repetitive DNA sequences of various Bifidobacterium species. Conclusions:, Ten of 37 experimental primer sets showed specificity for B. adolescentis, B. angulatum, B. pseudocatenulatum, B. breve, B. bifidum, B. longum, B. longum biovar infantis and B. dentium. Significance and Impact of the Study:, The results suggest that published Bifidobacterium primer sets should be re-evaluated for both reproducibility and specificity for the identification of Bifidobacterium species using PCR. Improvement of existing PCR methods will be needed to facilitate identification of other Bifidobacterium strains, such as B. animalis, B. catenulatum, B. thermophilum and B. subtile. [source]

Development of microsatellite DNA markers and their chromosome assignment in the common marmoset

AMERICAN JOURNAL OF PRIMATOLOGY, Issue 11 2009
Hideki Katoh
Abstract This study was performed to develop microsatellite DNA markers, which are useful for linkage analyses, gene mapping and blood chimera analyses in the common marmoset (Callithrix jacchus). We searched 153 of 295 bacterial artificial clone DNA sequences of the common marmoset that were archived in the NCBI database in 2004. On the basis of the search, we designed 186 PCR primer sets. When tested using 5 unrelated individuals, we successfully detected 154 markers with PCR products, of which 80 (52%) were polymorphic and 74 (48%) were monomorphic. We assigned each of the 154 markers to a human chromosome based on BLAST searches, which was achieved by searching the entire human genome sequences using an ,3,kb section of each forward primer sequence, including ,1.5,kb of the upstream and ,1.5,kb of the downstream sequences. Combining our assignment data and the chromosome painting-assisted karyotype of the common marmoset [Sherlock et al., Genomics 33:214,219, 1996], we prepared a list of 154 microsatellite DNA markers that were assigned to human chromosomes, except for the Y chromosome, which is equivalent to a chromosome map. Using five microsatellite DNA markers, we have established a fragment analysis method with a sequencer, which can be routinely used for blood chimera analysis, parentage diagnosis and individual identification. Am. J. Primatol. 71:912,918, 2009. © 2009 Wiley-Liss, Inc. [source]

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]