Search results for "Primer"
showing 10 items of 530 documents
Recombinant GII.P16 genotype challenges RT-PCR-based typing in region A of norovirus genome
2021
Abstract Objectives In latest years GII.4[P16] and GII.2[P16] noroviruses have become predominant in some temporal/geographical settings. In parallel with the emergence of the GII.P16 polymerase type, norovirus surveillance activity in Italy experienced increasing difficulties in generating sequence data on the RNA polymerase genomic region A, using the widely adopted JV12A/JV13B primer set. Two sets of modified primers (Deg1 and Deg2) were tested in order to improve amplification and typing of the polymerase gene. Methods Amplification and typing performance of region A primers was assessed in RT-PCR on 452 GII norovirus positive samples obtained from 2194 stool samples collected in 2016–2…
A practical method for barcoding and size-trimming PCR templates for amplicon sequencing
2016
Sample barcoding facilitates the analysis of tens or even hundreds of samples in a single next-generation sequencing (NGS) run, but more efficient methods are needed for high-throughput barcoding and size-trimming of long PCR products. Here we present a two-step PCR approach for barcoding followed by pool shearing, adapter ligation, and 5′ end selection for trimming sets of DNA templates of any size. Our new trimming method offers clear benefits for phylogenetic studies, since targeting exactly the same region maximizes the alignment and enables the use of operational taxonomic unit (OTU)-based algorithms.
High-Throughput Mapping of 2′-O-Me Residues in RNA Using Next-Generation Sequencing (Illumina RiboMethSeq Protocol)
2017
Detection of RNA modifications in native RNAs is a tedious and laborious task, since the global level of these residues is low and most of the suitable physico-chemical methods require purification of the RNA of interest almost to homogeneity. To overcome these limitations, methods based on RT-driven primer extension have been developed and successfully used, sometimes in combination with a specific chemical treatment. Nowadays, some of these approaches have been coupled to high-throughput sequencing technologies, allowing the access to transcriptome-wide data. RNA 2'-O-methylation is one of the ubiquitous nucleotide modifications found in many RNA types from bacteria, archaea, and eukarya.…
Association of liver steatosis with lipid oversecretion and hypotriglyceridaemia in C57BL/6j mice fed trans-10, cis-12-linoleic acid
2003
AbstractConjugated linoleic acids (CLA) have recently been recognized to reduce body fat and plasma lipids in some animals. This study demonstrated that the steatosis accompanying the fat loss induced by trans-10,cis-12-C18:2 (CLA2) and not cis-9,trans-11-C18:2 (CLA1) isomer in C57BL/6j mice was not due to an alteration of the liver lipoprotein production that was even increased. The 3-fold decrease in plasma triacylglycerol contents and the induction of mRNA expression of low-density lipoprotein receptors concomitantly observed in CLA2-fed mice suggested an increase in the lipoprotein clearance at the level of the liver itself. CLA1 feeding produced similar but attenuated effects on trigly…
Validation of a questionnaire of attributions to the detection of coincidences in academic work
2019
El plagio como tema de investigación en el contexto universitario no ha cesado de ser una preocupación constante debido a sus múltiples repercusiones. El presente estudio describe el proceso seguido en la validación del Cuestionario de atribuciones para la detección de coincidencias en trabajos académicos (CUDECO), cuya finalidad es evaluar la relevancia que tiene citar correctamente y detectar las causas que llevan a cometer plagio. A partir de una muestra de 2,331 estudiantes, se realiza un anális is de la fiabilidad como consistencia interna y un análisis factorial exploratorio (AFE) del instrumento. El análisis de la fiabilidad lleva a proponer un reajuste en el cuestionario inicialment…
Primera persona del plural en los juicios orales. Valor representativo y estrategia atenuante
2018
El objetivo de este artículo es doble, pues nos proponemos estudiar los valores que puede desarrollar la primera persona del plural en los juicios orales y, en relación con ello, examinar si es posible identificar usos atenuantes asociados a este mecanismo referencial. Tras analizar nuestro corpus siguiendo una metodología cualitativa y cuantitativa, hemos podido observar que el uso de la primera persona del plural en este género no solo se debe al carácter representativo del sistema legal español o a las imposiciones de la tradición discursiva. Frecuentemente, el uso de la primera persona del plural (especialmente el plural de modestia y el plural sociativo) codifica un valor atenuante ori…
Stereochemical features of the hydrolysis of 9,10-epoxystearic acid catalysed by plant and mammalian epoxide hydrolases
2002
cis-9,10-Epoxystearic acid was used as a tool to probe the active sites of epoxide hydrolases (EHs) of mammalian and plant origin. We have compared the stereochemical features of the hydrolysis of this substrate catalysed by soluble and membrane-bound rat liver EHs, by soluble EH (purified to apparent homogeneity) obtained from maize seedlings or celeriac roots, and by recombinant soybean EH expressed in yeast. Plant EHs were found to differ in their enantioselectivity, i.e. their ability to discriminate between the two enantiomers of 9,10-epoxystearic acid. For example, while the maize enzyme hydrated both enantiomers at the same rate, the EH from soybean exhibited very high enantioselecti…
Transcriptional Response of Saccharomyces cerevisiae to Different Nitrogen Concentrations during Alcoholic Fermentation▿ †
2007
Gene expression profiles of a wine strain of Saccharomyces cerevisiae PYCC4072 were monitored during alcoholic fermentations with three different nitrogen supplies: (i) control fermentation (with enough nitrogen to complete sugar fermentation), (ii) nitrogen-limiting fermentation, and (iii) the addition of nitrogen to the nitrogen-limiting fermentation (refed fermentation). Approximately 70% of the yeast transcriptome was altered in at least one of the fermentation stages studied, revealing the continuous adjustment of yeast cells to stressful conditions. Nitrogen concentration had a decisive effect on gene expression during fermentation. The largest changes in transcription profiles were o…
Primeras consideraciones a la Ley Orga?nica 3/2021, de 24 de marzo, de regulación de la eutanasia. el reconocimiento de un nuevo derecho
2021
The aim of this paper is to carry out a brief study of the recent Organic Law 3/2021, of March 24, on the regulation of euthanasia (BOE. March 25, 2021) approved in Spain, from a constitutional perspective, highlighting its main aspects. The matter under study is not exempt from controversy because it covers legal, medical and ethical considerations of great complexity.
Fibronectin-binding nanoparticles for intracellular targeting addressed by B. burgdorferi BBK32 protein fragments.
2011
Virus-like particles (VLPs) are created by the self-assembly of multiple copies of envelope and/or capsid proteins from many viruses, mimicking the conformation of a native virus. Such noninfectious nanostructures are mainly used as antigen-presenting platforms, especially in vaccine research; however, some of them recently were used as scaffolds in biotechnology to produce targeted nanoparticles for intracellular delivery. This study demonstrates the creation of fusion VLPs using hepatitis B core protein-based system maintaining a fibronectin-binding property from B. burgdorferi BBK32 protein, including the evidence of particles’ transmission to BHK-21 target cells via caveolae/rafts endoc…