comparison of microarray and quantitative real time pcr methods for measuring microrna levels in msc cultures
Báo cáo khoa học:" Development of TaqMan® MGB fluorescent real-time PCR assay for the detection of anatid herpesvirus 1?" pdf
... containing a wide range of viral DNA amounts, eliminating the need for sample dilution and minimizing sample handling The results for intra- and inter-assay precision indicate that both intra-assay ... Establishment of the fluorescent quantitative real- time PCR standard curve Standard curve of the AHV-1 fluorescent quantitative real- time PCR Ten-fold dilutions of standard DNA ranging from 1.0 ... improvement of the previously developed ordinary TaqMan real- time PCR method Results Development and optimization of FQ -PCR and conventional PCR Following the optimization of FQ -PCR, final concentrations...
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Báo cáo khoa học: " Development of TaqMan® MGB fluorescent real-time PCR assay for the detection of anatid herpesvirus 1" pdf
... containing a wide range of viral DNA amounts, eliminating the need for sample dilution and minimizing sample handling The results for intra- and inter-assay precision indicate that both intra-assay ... Establishment of the fluorescent quantitative real- time PCR standard curve Standard curve of the AHV-1 fluorescent quantitative real- time PCR Ten-fold dilutions of standard DNA ranging from 1.0 ... improvement of the previously developed ordinary TaqMan real- time PCR method Results Development and optimization of FQ -PCR and conventional PCR Following the optimization of FQ -PCR, final concentrations...
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Tài liệu Comparison of lung cancer cell lines representing four histopathological subtypes with gene expression profiling using quantitative real-time PCR pptx
... patients and lung cancer cells [1-3,6-10], marker gene candidates have varied depending on the report Quantitative real- time PCR (qPCR) is generally considered the “gold-standard” assay for measuring ... 1) In immunostained tumor tissues, AD cells showed immunostaining of S100P in the cytoplasm and the nucleus, while SQ cells showed immunostaining of RAB25 in the cytoplasm The localization of ... Experiments (cell culture and DNA microarray analysis) YD: Experiments (cell culture and quantitative real- time PCR analysis) YF: Experiments (cell culture and quantitative real- time PCR analysis) MI:...
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Báo cáo sinh học: " Determination of suitable housekeeping genes for normalisation of quantitative real time PCR analysis of cells infected with human immunodeficiency virus and herpes viruses" pdf
... determining the suitability of 10 commonly used reference genes in the context of infection with various strains of HIV-1, HSV-1, CMV and VZV in a range of both primary and cultured cell lines using ... strains Towne and Toledo, VZV strain Schenke and HIV-1 strains BaL, NL4.3 and RFW were propagated according to standard protocols [16-20] The cell types infected, MOI, time of infection and percentage ... (NC-1 strain), CMV (Toledo and Towne strains) and VZV (Schenke strain) and harvested at the earliest time of maximal productive infection as illustrated in Table The expression levels of 10 commonly...
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Báo cáo hóa học: " Determination of suitable housekeeping genes for normalisation of quantitative real time PCR analysis of cells infected with human immunodeficiency virus and herpes viruses" pot
... determining the suitability of 10 commonly used reference genes in the context of infection with various strains of HIV-1, HSV-1, CMV and VZV in a range of both primary and cultured cell lines using ... strains Towne and Toledo, VZV strain Schenke and HIV-1 strains BaL, NL4.3 and RFW were propagated according to standard protocols [16-20] The cell types infected, MOI, time of infection and percentage ... (NC-1 strain), CMV (Toledo and Towne strains) and VZV (Schenke strain) and harvested at the earliest time of maximal productive infection as illustrated in Table The expression levels of 10 commonly...
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báo cáo khoa học: " Selection of reference genes for quantitative real-time PCR expression studies in the apomictic and sexual grass Brachiaria brizantha" ppt
... run in a BioRad qRT -PCR machine using the following cycling parameters: 94C for min, 40 cycles of 94C for 15 s, 60C for 10 s, 72C for 15 s and 60C for 35 s No-template controls (NTC) were included ... and sexual reproduction within Brachiaria makes it an interesting system for understanding the molecular pathways involved in both modes of reproduction The identification of genes involved in ... normalization of Real- Time quantitative RT -PCR by geometric averaging of multiple internal control genes Genome Biol 2002, 3:34 Brunner AM, Yakovlev IA, Strauss SH: Validating internal controls for quantitative...
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báo cáo khoa học: " Validation of reference genes for quantitative real-time PCR during leaf and flower development in Petunia hybrida" pot
... both lines In contrast, the suggested genes did not coincide and were EF1a and SAND in Mitchell, whilst CYP and RAN1 were the genes of choice in V30 In conclusion, we provide a list of genes in ... μL of cDNA, mM MgCl2, 0.2 mM each dNTP, 0.4 μL of each primer and 1.25 U enzyme Real- time PCR Mitchell Real- time PCR was performed in an Mx 3005P QPCR system (Stratagene, La Jolla, CA) using ... for normalization was the same for both lines: using the proposed cut-off of 0.15 and comparing single developmental stages, the required number was two for Mitchell and V30 The requirement for...
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Báo cáo khoa học: "Detection of anatid herpesvirus 1 gC gene by TaqMan™ fluorescent quantitative real-time PCR with specific primers and probe" pps
... detect low-level of viral loads FQ -PCR is based on the conventional principles of PCR and has being become an increasingly popular way for the diagnosis of bacteria and viruses infection The diagnostic ... to further investigate pcDNA3.1-gC DNA vaccine in vivo in ducks Results Development and optimization of a TaqMan™ FQ -PCR Final concentrations of primers each of 0.5 μmol/L and probe of 0.25 μmol/L ... poor performance in quantitation and a relative waste of time It is not suitable for large-scale applications In recent times, a more sensitive, time- saving and advanced method has emerged in the...
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Báo cáo sinh học: " Reference gene selection for quantitative real-time PCR analysis in virus infected cells: SARS corona virus, Yellow fever virus, Human Herpesvirus-6, Camelpox virus and Cytomegalovirus infections" doc
... The respective MOI and time of cell culture are shown in table and were chosen to allow maximal infection as determined by immunofluorescence and real- time PCR [8-11] For kinetic studies, cells ... BestKeeper indices; even actin was included into our gene panel These findings demonstrate the usefulness of analysing a wide variety of reference gene candidates The inconsistent data regarding to ... conditions and results of virus kinetics CMV CAMP SARS YF MRC-5 2.0 72 100 0,6,12,24,48,72 cell line multiplicity of infection time to maximal infection /h max infected cells % measuring point /h...
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báo cáo hóa học:" Reference gene selection for quantitative real-time PCR analysis in virus infected cells: SARS corona virus, Yellow fever virus, Human Herpesvirus-6, Camelpox virus and Cytomegalovirus infections" doc
... The respective MOI and time of cell culture are shown in table and were chosen to allow maximal infection as determined by immunofluorescence and real- time PCR [8-11] For kinetic studies, cells ... BestKeeper indices; even actin was included into our gene panel These findings demonstrate the usefulness of analysing a wide variety of reference gene candidates The inconsistent data regarding to ... conditions and results of virus kinetics CMV CAMP SARS YF MRC-5 2.0 72 100 0,6,12,24,48,72 cell line multiplicity of infection time to maximal infection /h max infected cells % measuring point /h...
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Báo cáo khoa học: " Establishment of one-step SYBR green-based real time-PCR assay for rapid detection and quantification of chikungunya virus infection" pps
... tool for rapid detection and quantification of CHIKV during natural infection, in research laboratory settings and possibly monitoring the extent of viral replication in patients for clinical ... Health Laboratory, Ministry of Health, Singapore) CHIKV was propagated in Vero and C6/36 cells and infectious virus titers were determined by plaque forming assay using BHK cells in the Biosafety ... (GenBank accession no: AF490259 of Ross reference strain, EU703762.1 of Malaysia strain, EF027140.1 of Indian strain, AM258995.1 of Reunion strain and EF452494.1 of USA strain), the potential target...
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Báo cáo sinh học: "Quantitative real-time PCR study on persistence of pDNA vaccine pVax-Hsp60 TM814 in beef muscles" pptx
... approve usage of DNA vaccines in food- producing animals In this work, the QRTPCR method was used for the study of the persistence of pDNA at the injection sites in mice and beef cattle For this reason ... muscle (at the injection site) after administration of 10 μg pDNAX in 5-week Levels of pDNAX detected by QRTPCR in calf muscle (at the injection site) after administration of 10 μg pDNAX in 5-week ... fluorescent liposomes and analysis of gene expression Single dose of pLacZ (10 μg) was injected into calf muscle of BALB/c mice (5 weeks of age) Plasmid pLacZ was delivered in the following forms: naked...
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Báo cáo hóa học: " A quantitative real time PCR method to analyze T cell receptor Vb subgroup expansion by staphylococcal superantigens" doc
... phospholipase C and phosphokinase C pathways [10], leading to a massive production of proinflammatory cytokines including interleukin-2 and interferon-g [11], resulting in extensive proliferation of T cells ... Primers primers used in qRT -PCR have uniform and high efficiency and linearity The specificity of qRT -PCR using SYBR Green I platform was often determined by analyzing melting curves In this study, ... specificities of each primer set were determined by analyzing melting curves and sequencing amplified PCR products Melting curve analysis and sequencing amplified PCR products of reactions for, Cb and...
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Báo cáo khoa học: " Development of a fluorescent quantitative real-time polymerase chain reaction assay for the detection of Goose parvovirus in vivo" ppsx
... Establishment of the fluorescent quantitative real- time PCR (FQ -PCR) standard curve Establishment of the fluorescent quantitative real- time PCR (FQ -PCR) standard curve Ten-fold dilutions of standard ... sites of GPV invasion in infected goslings Methods Virus and PCR template DNA preparation GPV CHV strain, a high-virulence strain of GPV, was obtained from Key Laboratory of Animal Diseases and ... PCR and FQ -PCR protocol The detection limit of the conventional PCR was determined based on the Fifty goslings were randomly divided into groups In brief, a group of 40 goslings were orally infected...
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Báo cáo y học: "Endothelial Cytomegalovirus infection monitored by quantitative real-time PCR in critically ill patients" pps
... Bordes et al Critical Care 2011, 15:412 http://ccforum.com/content/15/2/412 Page of Figure Cytomegalovirus plasma load measurements during ICU stay of four severe burn patients Table Patient characteristics ... 30 9,130 Discharged from ICU 141 63,400 Died in ICU 133 130,000 Discharged from ICU 105 137,000 Died in ICU ICU stay (days) Viremia peak is expressed in copies/ml DBSA, deep burn surface area;...
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Quantitative real time PCR
... Huggett, and Stephen Bustin Abstract The MIQE (minimum information for the publication of quantitative real- time PCR) guidelines were published in 2009 with the twin aims of providing a blueprint for ... expression Introduction The MIQE (minimum information for the publication of quantitative real- time PCR) guidelines [1] represent a major milestone in the transformation of the real- time quantitative ... used in real- time quantitative PCR (qPCR) dPCR involves partitioning a single PCR reaction into hundreds or thousands of subreactions under conditions where some of the subreactions amplify, indicating...
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Báo cáo khoa học: "Comparison of conformal and intensity modulated radiation therapy techniques for treatment of pelvic tumors. Analysis of acute toxicity" pdf
... “increased frequency or change in quality of bowel habits nor requiring medication or rectal discomfort not requiring analgesics” [27], this finding is not relevant in the clinical practice and ... very uncomfortable symptom and can cause dehydration and malabsorption of vitamins, lactose, and bile acids [4] Another important finding was the higher absence of GI symptoms (grade 0) in IMRT ... in clinical practice and definition of this grade could be subjective, as the information collected was based on physician’s notes in patient’s charts Our results of lower acute GI toxicity in...
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Báo cáo sinh học: " Comparison of real-time PCR and hemagglutination assay for quantitation of human polyomavirus JC" pptx
... Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral load A – C: Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral ... [21,26] We first examined the fidelity of real- time quantitative PCR in detecting and quantitating JCV T-antigen gene sequences from clinical specimens Quantitative real- time PCR proved to be an ... dynamic range, and real- time PCR was more reliable than the HA assay for in vitro calculation of initial virus inoculum and replicated virus In the future, quantitative real- time PCR can be employed...
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báo cáo hóa học:"Comparison of real-time PCR and hemagglutination assay for quantitation of human polyomavirus JC" potx
... Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral load A – C: Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral ... [21,26] We first examined the fidelity of real- time quantitative PCR in detecting and quantitating JCV T-antigen gene sequences from clinical specimens Quantitative real- time PCR proved to be an ... dynamic range, and real- time PCR was more reliable than the HA assay for in vitro calculation of initial virus inoculum and replicated virus In the future, quantitative real- time PCR can be employed...
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Báo cáo sinh học: " Detection, quantification and genotyping of Herpes Simplex Virus in cervicovaginal secretions by real-time PCR: a cross sectional survey" potx
... questionnaire and an examination for genital lesion by the clinician/nursing officer The study was approved and Detection of HSV DNA, probe typing & melting point determination Quantitative PCR was performed ... having Bacterial Vaginosis (BV) Twenty-nine of 70 (41%) subjects had Candida and Trichomonas vaginalis was found in out of 70 (10%) Eight out of 62 (13%) were positive for Hepatitis B and out of ... typing in a single step This may have circumvented problems relating to sensitivity of the HSV-1 and -2 probe relative binding In a single step multiplex assay mutations in the probe binding...
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