Search results for "PCR"

Tyramine and phenylethylamine production among lactic acid bacteria isolated from wine.

2007

The ability of wine lactic acid bacteria to produce tyramine and phenylethylamine was investigated by biochemical and genetic methods. An easy and accurate plate medium was developed to detect tyramine-producer strains, and a specific PCR assay that detects the presence of tdc gene was employed. All strains possessing the tdc gene were shown to produce tyramine and phenylethylamine. Wines containing high quantities of tyramine and phenylethylamine were found to contain Lactobacillus brevis or Lactobacillus hilgardii. The main tyramine producer was L. brevis. The ability to produce tyramine was absent or infrequent in the rest of the analysed wine species.

Quantification of PCR products by phosphate measurement

2008

Various techniques for quantification of PCR are available. Most frequently, the densitometric intensities of ethidium bromide-stained PCR products separated in gels are compared after normalizing to the levels of housekeeping gene products such as beta-actin. More precise, but extremely time consuming, is the technique of competitive PCR. Newer methods, such as tracking amplification in real-time, have high start-up and maintenance costs (e.g., TaqMan, Applied Biosystems; LightCycler, Roche; I-Cycler, Bio-Rad). Here, I describe an alternative, simple technique to quantify PCR products by determining the entire phosphate released during PCR. The method can be performed using common laborato…

Exploring the mechanisms by which reindeer droppings induce fen peat methane production

2021

Abstract Peatlands, especially fens, are known to emit methane. Reindeer (Rangifer tarandus) use mires mainly as spring and summer pastures. In this work we observed that adding reindeer droppings to fen peat increased the potential methane production by 40%. This became apparent when droppings originating from reindeer kept in pen or pasture in winter were added to methanogenic fen peat samples. The droppings introduced Methanobacteriaceae (Methanobrevibacter; > 90% of the mcrA MiSeq reads) to the peat, which was originally populated by Methanosarcinaceae, Methanosaetaceae, Methanoregulaceae, Methanobacteriaceae, Methanomassiliicoccaceae, Methanocellaceae and Methanomicrobiaceae. The origi…

Molecular diagnostic tools for the detection and characterization of Phoma tracheiphila

2011

In the recent years PCR-based techniques for the identification and detection of Phoma tracheiphila, the causal agent of citrus mal secco disease, have been evaluated aiming to provide tools for biological and epidemiological studies. A wide collection of P. tracheiphila strains was used to evaluate and validate diagnostic protocols and a fAFLP method for fungal characterization. Conventional and real-time PCR protocols were successfully tested for the specific identification of P. tracheiphila and its detection in planta. A further improvement of the real-time PCR protocol and the DNA extraction methods allowed the quantification of the fungus both from naturally infected and artificially …

Does a specific genetic background influence endurance or power-orientated phenotypes?

2012

The purpose of this study was to determine the probability of individuals having the “best” mitochondrial biogenesis related polygenic profile that could increase performance. We compared polygenic profile analyzing several polymorphisms on sixty professional italian soccer players, considered “power-oriented athletes” and thirty sedentary volunteers. Samples of venous blood were obtained by standard clinical procedures and anticoagulant-treated blood was used to prepare genomic DNA. The polymorphic sites were scanned using PCR-RFLP protocols with different enzyme. Furthermore, a cloted part of venus blood sample was used to obtain serum from which we analyzed total cholesterol, high densit…

Polymerase Chain Reaction (PCR)

2008

Effect of transfection with PLP2 antisense oligonucleotides on gene expression of cadmium-treated MDA-MB231 breast cancer cells

2012

Emerging evidence indicates that cadmium (Cd) is able to regulate gene expression, drastically affecting the pattern of transcriptional activity in human normal and pathological cells. We have already shown that exposure of MDA-MB231 breast cancer cells to 5 μM CdCl(2) for 96 h, apart from significantly affecting mitochondrial metabolism, induces modifications of the expression level of genes coding for members of stress response-, mitochondrial respiration-, MAP kinase-, NF-κB-, and apoptosis-related pathways. In the present study, we have expanded the knowledge on the biological effects of Cd-breast cancer cell interactions, indicating PLP2 (proteolipid protein-2) as a novel member of the…

Retrospective screening for SARS-CoV-2 among influenza-like illness hospitalizations: 2018-2019 and 2019-2020 seasons, Valencia region, Spain

2021

Este artículo se encuentra disponible en la siguiente URL: https://onlinelibrary.wiley.com/doi/epdf/10.1111/irv.12899 En este artículo de investigación también participan: Juan Mollar-Maseres, Germán Schwarz-Chavarri, Sandra García-Esteban, Joan Puig-Barberà, Javier Díez-Domingo y F. Xavier López-Labrador. On 9 March 2020, the World Health Organization (WHO) Global Influenza Programme (GIP) asked participant sites on the Global Influenza Hospital Surveillance Network (GIHSN) to contribute to data collection concerning severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We re-analysed 5833 viral RNA archived samples collected prospectively from hospital admissions for influenza-lik…

Quality controls for cell cultures: identification of interspecies cross-contamination by PCR-RFLP analysis of the cytochrome b gene

2012

Cross-contaminations of a cell line with cells of different species represent a potential risk in laboratories handling human and animal cells. Therefore, it is necessary to control such contaminations. Tests based on mitochondrial DNA (mtDNA) are used in forensic analysis, phylogenetic studies and in food authentication. However, the use of mtDNA in quality controls of cell cultures is recent. Mitochondrial sequence differences of closely related animal species are five- to tenfold higher than those of nuclear genes. On the contrary, intraspecies variation in mitochondrial sequences is low in most animal species. Moreover, each cell contains 100–10.000 mitochondrial genomes. The amount of …

Study of variability of an endophytic acremonium population in symptomless grapevine

2009

The endophytism of genus Acremonium in symptomless grapevines of several cultivars, grown both in pots or in the field, is reported. The no-tillage management of tested grapevines and the lack of chemical treatments facilitated the investigation of the ecological plant-fungus interaction. The isolates were differentiated at the strain level by vegetative compatibility assay and randomly amplified polymorphic DNA (RAPD) analyses, using a set of four oligo-nucleotide primers. The endophytic Acremonium population in the healthy grapevine object of the survey seemed to show a low variability; all the techniques agreed in grouping the Acremonium strains in three different clusters and no relatio…