Search results for " PCR"

Droplet digital PCR as a tool for investigating dynamics of cryptic symbionts

2021

Abstract Interactions among symbiotic organisms and their hosts are major drivers of ecological and evolutionary processes. Monitoring the infection patterns among natural populations and identifying factors affecting these interactions are critical for understanding symbiont–host relationships. However, many of these interactions remain understudied since the knowledge about the symbiont species is lacking, which hinders the development of appropriate tools. In this study, we developed a digital droplet PCR (ddPCR) assay based on apicomplexan COX1 gene to detect an undescribed agamococcidian symbiont. We show that the method gives precise and reproducible results and enables detecting cryp…

Becton Dickinson Directigen EZ Flu A+B assay in the diagnosis of pandemic influenza A H1N1 2009 virus infection in adult patients

2011

Validation of a set of reference genes to study response to herbicide stress in grasses

2012

Abstract Background Non-target-site based resistance to herbicides is a major threat to the chemical control of agronomically noxious weeds. This adaptive trait is endowed by differences in the expression of a number of genes in plants that are resistant or sensitive to herbicides. Quantification of the expression of such genes requires normalising qPCR data using reference genes with stable expression in the system studied as internal standards. The aim of this study was to validate reference genes in Alopecurus myosuroides, a grass (Poaceae) weed of economic and agronomic importance with no genomic resources. Results The stability of 11 candidate reference genes was assessed in plants res…

DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen

2020

Droplet microfluidics is a technology that enables the production and manipulation of small volumes. In biosciences, the most popular application of this technology is Droplet Digital™ PCR (ddPCR™), where parallel nanoliter-scale PCR assays are used to provide a high sensitivity and specificity for DNA detection. However, the recovery of PCR products for downstream applications such as sequencing can be challenging due to the droplets' stability. Here we compared five methods for disrupting the droplets to recover DNA. We found that rapid freezing in liquid nitrogen results in a clear phase separation and recovery of up to 70% of the DNA content. Liquid nitrogen freezing can thus offer a si…

Differential expression of two glucocorticoid receptors in seabass (teleost fish) head kidney after exogeneous cortisol inoculation

2009

Stressful conditions include a prompt release of corticosteroid hormones which can mediate gene expression through glucocorticoid receptors (GR). Since two seabass (Dicentrarchus labrax) GRs have been cloned and sequenced from peritoneal cavity cells (DlGR1) and liver (DlGR2), a comparative amino acid sequence analysis that included Haplochromis burtoni HbGRs, was carried out and homologies disclosed. The DlGR1 and DlGR2 deduced aminoacid sequences showed 61% identity (I) and 70% similarity (S). Moreover, DlGR2 was similar to HbGR2b (69% I, 73% S), and the DlGR1 to HbGR1 (72% I, 78% S). In addition, we examined the expression of the DlGRs after exogeneous cortisol inoculation into the perit…

Glucose transporter 4 mRNA expression in subcutaneous adipose tissue of women with PCOS remains unchanged despite metformin withdrawal: is there a ce…

2021

Purpose: Metformin induces GLUT-4 mRNA expression in insulin target tissues in PCOS. It is unclear how long this impact is sustained after withdrawal of metformin. We aimed to compare the effect of metformin withdrawal on GLUT-4 mRNA expression in subcutaneous adipose tissue after prior short (ST, 1 year, N = 11) and long term (LT, at least 3 years, N = 13) treatment in obese PCOS women. Methods: At baseline and 6 months after withdrawal, biopsy of subcutaneous adipose tissue followed by quantitative PCR analysis was performed to determine GLUT-4 mRNA expression. Results: We found no time/effect differences in GLUT-4 mRNA expression in ST (2-dCt at baseline 0.42 (0.16–0.48) vs 2-dCt after 6…

Differences in cytomegalovirus plasma viral loads measured in allogeneic hematopoietic stem cell transplant recipients using two commercial real-time…

2010

5 páginas, 3 figuras.

Prime indicazioni sulle biocenosi associate alle mummie delle Catacombe dei Cappuccini di Palermo

2009

La fama del convento, costruito dai Cappuccini di Palermo nel 1533, è dovuta in gran parte alle famose "Catacombe". L’origine delle catacombe si fa risalire al 1599 circa, quando i frati, sfruttando una preesistente cavità naturale al di sotto dell’altare maggiore della chiesa, trasferirono le salme di 40 frati precedentemente sepolti presso il lato meridionale della chiesa stessa. I frati iniziarono ad ampliare le catacombe, scavando nel sottosuolo, in relazione al crescente numero di nuove salme che nel tempo si conservavano. Allo stato attuale le catacombe sono una sorta di cimitero sotterraneo, in uso nel XVII secolo, costituito da lunghe gallerie scavate nel tufo per un’estensione di c…

EVALUATING THE RISK OF HEPATITIS B REACTIVATION IN PATIENTS WITH HAEMATOLOGICAL MALIGNANCIES: IS THE SERUM HEPATITIS B PROFILE RELIABLE?

2009

Background/Aim: Patients with an occult hepatitis B virus (HBV) infection undergoing deep immunosuppression are potentially at risk of HBV reactivation. In order to assess whether a polymerase chain reaction (PCR) assay for HBV DNA in serum could be used to predict the reactivation of an occult HBV infection, we performed a retrospective study in a cohort of Sicilian patients with oncohaematological diseases. Methods: We studied by a highly sensitive ad hoc nested PCR for serum HBV DNA 75 HBsAg-negative oncohaematological patients requiring chemotherapy. Results: Thirty-three patients (44%) were HBV seronegative (anti-HBc and anti-HBs negative) and 42 patients (56%) were HBV seropositive (a…

Metodo per la indicizzazione approssimata e il reperimento di sequenze che si ripetono all'interno di sequenze

2005

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