quantitative real time pcr in adna research

... 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; ... 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 Patient Age (years) TBSA...

Ngày tải lên: 14/08/2014, 07:21

... ranked actin at the last position, indicating that it is an unsuitable reference gene in virus infected cells The actin gene shows significant variations with increasing degree of infection The ... and virus detection by real- time PCR Camelpox strain CP-19, CMV strain AD169, HHV-6 strain U1102, SARS coronavirus strain 6109 and YFV strain 17D were propagated according to standard procedures ... data set according to the Bestkeeper tool revealed very good BestKeeper indices; even actin was included into our gene panel These findings demonstrate the usefulness of analysing a wide variety...

Ngày tải lên: 18/06/2014, 22:20

... ranked actin at the last position, indicating that it is an unsuitable reference gene in virus infected cells The actin gene shows significant variations with increasing degree of infection The ... and virus detection by real- time PCR Camelpox strain CP-19, CMV strain AD169, HHV-6 strain U1102, SARS coronavirus strain 6109 and YFV strain 17D were propagated according to standard procedures ... data set according to the Bestkeeper tool revealed very good BestKeeper indices; even actin was included into our gene panel These findings demonstrate the usefulness of analysing a wide variety...

Ngày tải lên: 20/06/2014, 04:20

... agarose gel electrophoresis Real- time PCR conditions and analysis PCR reactions were performed in 96-well plates with the Chromo4 Real- Time PCR Detector System (BioRadđ) using SYBRđ Green to detect ... real- time PCR analysis, and three technical replicates were analyzed for each biological replicate Reactions were run in a BioRad qRT -PCR machine using the following cycling parameters: 94C for min, ... genes for quantitative real- time PCR in bovine preimplantation embryos BMC Dev Biol 2005, 5:27 Mamo S, Gal A, Bodo S, Dinnyes A: Quantitative evaluation and selection of reference genes in mouse...

Ngày tải lên: 12/08/2014, 03:20

... 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 a SYBR Green based PCR assay (with ROX as the optional ... developmental stages in Mitchell and V30 Real- time PCR reactions were performed on the six cDNA samples obtained from each Petunia line with the nine primer pairs representing the candidate RG In order to ... normalization or routinely used as controls for northern blots or RT -PCR From the original list we developed a short list of nine, including genes encoding Actin-11 (ACT), Cyclophilin-2 (CYP) [10],...

Ngày tải lên: 12/08/2014, 03:21

... of DNA vaccinated livestock, mainly in muscles Nowadays, there are no definitive guidelines available to approve usage of DNA vaccines in food- producing animals In this work, the QRTPCR method ... hsp60 gene, generating 161 bp specific product 500 ng of genomic DNA (gDNA) template was amplified in duplicate in glass capillaries in a final volume of 20 μl using 2× Real time PCR Syber green ... detected by QRTPCR in calf 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...

Ngày tải lên: 14/08/2014, 19:22

... Furihata C: Dose-dependent alterations in gene expression in mouse liver induced by diethylnitrosamine and ethylnitrosourea and determined by quantitative real- time PCR Mutat Res 2009, 673:9-20 doi:10.1186/1475-2867-10-2 ... Comparison of lung cancer cell lines representing four histopathological subtypes with gene expression profiling using quantitative real- time PCR Cancer Cell International 2010 10:2 Publish with ... calcium-binding protein P, may be a good biomarker for AD cells The expression ratio of S100P in cancer/normal cells was high in AD samples and low in SQ samples In our study, all three AD cell lines...

Ngày tải lên: 15/02/2014, 04:20

... 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 bearing ... determined by analyzing melting curves and sequencing amplified PCR products Melting curve analysis and sequencing amplified PCR products of reactions for, Cb and Vb subgroups consisting of a single ... 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,...

Ngày tải lên: 18/06/2014, 16:20

... RNase H (Invitrogen) and diluted 1:100 before use for QPCR Quantitative PCR In order to measure housekeeping gene expression, cDNA was subject to QPCR using the platinum QPCR super mix kit (Invitrogen) ... reflecting variability in both virus strain and cell type Notable differences included 18sRNA being a relatively reliable gene in MDDCs infected with HIVBaL, but unreliable in other HIV infections ... 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...

Ngày tải lên: 18/06/2014, 18:20

... RNase H (Invitrogen) and diluted 1:100 before use for QPCR Quantitative PCR In order to measure housekeeping gene expression, cDNA was subject to QPCR using the platinum QPCR super mix kit (Invitrogen) ... reflecting variability in both virus strain and cell type Notable differences included 18sRNA being a relatively reliable gene in MDDCs infected with HIVBaL, but unreliable in other HIV infections ... 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...

Ngày tải lên: 20/06/2014, 01:20

... time It is not suitable for large-scale applications In recent times, a more sensitive, time- saving and advanced method has emerged in the field, which is fluorescent quantitative real- time PCR ... organs in a short time by intramuscular injection of DNA vaccine [51,52] In this study, AHV-1 gC can be detected in all analyzed tissues at hour postinoculation, and the concentration of gC maintained ... of the FQ -PCR was determined based on the highest dilution that resulted in the presence of Ct value in real- time PCR detection The detection limit of the conventional PCR was determined through...

Ngày tải lên: 12/08/2014, 04:21

... publication of quantitative real- time PCR) guidelines were published in 2009 with the twin aims of providing a blueprint for good real- time quantitative polymerase chain reaction (qPCR) assay design ... 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 ... (minimum information for the publication of quantitative real- time PCR) guidelines [1] represent a major milestone in the transformation of the real- time quantitative polymerase chain reaction (qPCR)...

Ngày tải lên: 05/09/2015, 10:35

... of microcystin-degrading bacteria in water environment We estimated microcystin-degrading bacteria in a biofilm from a practical biological treatment facility by real- time PCR using the newly ... This bacterium is included in the genus Sphingomonas (accession number AB110635) The strain C-1 was isolated from Lake Hongfeng in China and this strain is included in the genus Sphingopyxis (AB161684) ... Co USA), made up to a final volume of 50 µL using pure water Real- time PCR was performed using a 7,500 real- time PCR system (Applied Biosystems, USA) The DNA of C-1 strain was used as a positive...

Ngày tải lên: 05/09/2013, 10:15

... Cell lines were serially diluted (10-fold) in × 107 to × 107 lymphocytes to give carcinoma cell: lymphocyte ratios ranging from 1:10 to 1:107 CEA mRNA Analysis by Real- Time Quantitative PCR Quantitative ... and peritoneal washing [15] of gastric cancer patients by using real- time PCR for CEA mRNA The goal of this study was to evaluate the effectiveness of the CEA mRNA real- time PCR technique for ... classified into PCR- based methods and cytometric methods [8] With the advent of quantitative real- time PCR techniques [9], precise quantification of a target sequence has become possible Quantitative PCR...

Ngày tải lên: 18/06/2014, 16:20

... specific binding assay HSV type was confirmed in samples in which an equivocal typing result was initially obtained HSV type was confirmed by repeating the realtime assay using the Rotorgene 3000 instrument ... were used in a single tube real time PCR using the QuantiTect probe kit (QIAGEN Ltd, Crawley, UK) with the following cycling conditions using either an Opticon (GRI/MJ Research, Braintree, UK) ... 'spiked' into a PCR reaction containing approximately 100 copies of bacteriophage λ DNA and a primer pair directed against λ DNA The performance of the PCR was monitored by quantitative real- time PCR...

Ngày tải lên: 19/06/2014, 08:20

... the real- time qPCR The performance of the new qPCR assay was examined by determining the sensitivity, specificity, inter- and intraassay variability HBV DNA standard was obtained by end point ... sero-status Real- time quantitative PCR for HBV DNA DNA was extracted from 200 µL of serum using the QIAamp DNA Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions ... the International HBV DNA standard Data management The data obtained in the ABi real time machine after the PCR amplification and quantification of DNA was exported as an Excel spreadsheet into...

Ngày tải lên: 20/06/2014, 01:20

... specific binding assay HSV type was confirmed in samples in which an equivocal typing result was initially obtained HSV type was confirmed by repeating the realtime assay using the Rotorgene 3000 instrument ... were used in a single tube real time PCR using the QuantiTect probe kit (QIAGEN Ltd, Crawley, UK) with the following cycling conditions using either an Opticon (GRI/MJ Research, Braintree, UK) ... 'spiked' into a PCR reaction containing approximately 100 copies of bacteriophage λ DNA and a primer pair directed against λ DNA The performance of the PCR was monitored by quantitative real- time PCR...

Ngày tải lên: 20/06/2014, 04:20

... implemented by binding the real- time IRQ and the real- time tasks to one or more “shielded” processors, which never handle nonrealtime IRQs or tasks Of course, the nonreal -time IRQs and nonreal -time tasks ... hard real- time if a real- time application catches its deadline the 99.999% of the times The indeterminism caused by the hardware cannot be reduced by the software, thus no real- time operating ... processes instead of the real- time process’ code The operating system latency is the time elapsed between the instant in which an event is raised by some hardware device and the instant in which a real- time...

Ngày tải lên: 22/06/2014, 19:20

... efficient in harvesting bacterial DNA and reducing the remaining PCR inhibitors Therefore, the QIAamp DNA Mini Kit was utilized for improved efficacy of the multiplex real- time PCR assay in this ... multiplex real- time PCR for Salmonella (S.) Typhimurium The results of the multiplex real- time PCR were determined using decimal dilution of S Typhimurium ATCC 14028 DNA The PCR reaction contained ... real- time PCR assays [8,27] These advantages improved the multiplex real- time PCR assay in terms of shortening turnaround time for bacterial detection and reducing the risk of crosscontamination during...

Ngày tải lên: 07/08/2014, 23:22

... comparing the change in the treated group with the change in the placebo control Use of Q -PCR using RA and OA synovia To determine whether the Q -PCR technique can detect differences in cytokine ... patients originally relied on protein analysis of fluid compartments Blood sampling is often included in clinical trials, and measurement of certain plasma constituents, such as C-reactive protein, provides ... cDNA were included The same preparation of PBMC cDNA was used in all experiments, thereby allowing comparison of Q -PCR data obtained in different runs Standard curves were generated by linear regression...

Ngày tải lên: 09/08/2014, 01:23