Quantitative real-time Polymerase Chain Reaction (RT-qPCR) has become a valuable molecular technique in biomedical research. The selection of suitable endogenous reference genes is necessary for normalization of target gene expression in RT-qPCR experiments. The aim of this study was to determine the suitability of each 18S rRNA and ACTB as internal control genes for normalization of RT-qPCR data in some human cell lines transfected with small interfering RNA (siRNA). Four cancer cell lines including MCF-7, T47D, MDA-MB-231 and Hela cells along with HEK293 representing an embryonic cell line were depleted of E2F6 using siRNA specific for E2F6 compared to negative control cells, which were transfected with siRNA not specific for any gene. Using RT-qPCR, Ct (threshold cycle) values of 18S and ACTB were determined in transfected cells and compared with control cells. In the selection of the above cell lines, 18S was identified as the most stably expressed reference gene than ACTB in gene knockdown experiments.
Genotypic detection of some Antibiotics Resistant genes by using polymerase chain reaction (PCR). (20) Isolates of Acinetobacter baumannii that showed resistance to (Ceftaxim, Cefotaxim, Cefepim and Imipenim) were selected. The results showed that 20 isolates of A. baumannii possess the bla-OXA23 like gene, and that all isolates possess this gene with a percentage (100%). With molecular weight 605 bp. The current study showed that A. baumannii isolates carry 100% bla-OXA51like gene when studied with (20) isolates that are resistant to antibiotics (Imipenim Ceftazidime, Cifepime, Cifexime) that belong to this group of β-lactame with molecular weight 382 bp. Gene exp
... Show MoreAbstract: E2F6 is a member of the E2F family of transcription factors involved in regulation of a wide variety of genes through both activation and repression. E2F6 has been reported as overexpressed in breast cancers but whether or not this is important for tumor development is unclear. We first checked E2F6 expression in tumor cDNAs and the protein level in a range of breast cancer cell lines. RNA interference-mediated depletion was then used to assess the importance of E2F6 expression in cell lines with regard to cell cycle profile using fluorescence-activated cell sorting and a cell survival assay using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The overexpression of E2F6 was confirmed in breast tumor cDNA samp
... Show MoreBackground: Chronic myelogenous leukemia is a malignant hematological disease of hematopoietic stem cells. It is difficult to adapt treatment to each patient's risk level because there are currently few clinical tests and no molecular diagnostics that may predict a patient's clock for the advancement of CML at the time of chronic phase diagnosis. Biomarkers that can differentiate people based on the outcome at diagnosis are needed for blast crisis prevention and response improvement. Objective: This study is an effort to exploit the SLC25A3 gene as a potential biomarker for CML. Methods: RT-qPCR was applied to assess the expression levels of the SLC25A3 gene. Results: In comparison to the mean ΔCt of the control group, which was found to b
... Show MoremiRNAs regulate protein abundance and control diverse aspects of cellular processes and biological functions in metabolic diseases, such as obesity and diabetes. Lethal-7(Let-7) miRNAs specifically target genes associated with diabetes and have a role in the regulation of peripheral glucose metabolism. The present study aimed to describe the gene expressions of the let-7a gene with the development of diabetes in Iraq and the difference in the expression of this gene in patients with diabetes and healthy individuals. The association between age and gender with the development of diabetes was studied in this study and the results were compared with those of healthy individuals in the group of control. Based on the obtained results, there was
... Show MoreThe current study was conducted in the period extending from November 2018 to October 2019 and designed as a case-control study and aimed to assess the seroprevalence of HCMV. However, a total number of 91serum specimens were collected to fulfill this purpose from females (71 breast cancer patients, and control group of 20 females) attending Al-Amal hospital for cancer management and Baghdad teaching hospital and the practical part was performed in College of Science, University of Baghdad. The study protocol was approved by the Ethics Committee at the Department of Biology (Reference: BEC/0220/0011). The immunological part for evaluation of seroprevalence of HCMV was accomplished by ELISA technique which revealed that anti-HCMV IgG was sco
... Show MoreThe study included 200 samples were collected from children under two years included (50 samples from each of Cerebrospinal fluid, Blood, Stool and Urine) from, (Central Children Hospital and Children's Protections Educational Hospital) The Iraqi Ministry of Health, the Department of Health Baghdad .the period from the first of 2015 September to the first of December 2015, Were obtained isolates bacterial subjected to the cultural, microscopic and biochemical examination and diagnosed to the species by using vitek2 system .The results showed there were contamination in 6.5% of clinical samples. The diagnosed colonies which gave pink color on the MacConkey agar, golden yellow color on the Trypton Soy agar and green color on t
... Show MoreFor the period from February 2014 till May 2014, one hundred and nine lactose fermenter clinical isolates from different samples (urine, stool, wound swab, blood, and sputum) were collected from Alyarmok, Alkadimiya, and Baghdad teaching hospitals at Baghdad governorate. Identification of all Klebsiella pneumoniae isolates were carried out depending on macroscopic, microscopic characterizations, conventional biochemical tests, and Api 20E system. Fifty-three (48.62%) isolates represented K. pneumoniae; however, 51.73% represented other bacteria. Susceptibility test was achieved to all fifty-three K. pneumoniae isolates using five antibiotic disks (Ceftazidime, Ceftriaxone, Cefotaxime, Imipenem, and Meropenem). Most of tested isolates (90
... Show MoreThe present work aimed to investigate the neuraminidase (nan1) gene expression in 32 different clinical isolates of Pseudomonas aeruginosa to explore the role of the enzyme in different types of infection and might give a better understanding of host cell-pathogens interaction. In addition, the effect of monosaccharide D-mannose on neuraminidase gene expression in eight isolates was studied by utilizing a reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results demonstrated that the highest expression of nan1 gene was in otitis samples (208,913.81) which were significantly higher than that from other infections (P < 0.01). While, the concentrations of gene copies obtained from urin
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