extraction production of cdna and quantitative real time pcr

... primed cDNA- synthesis cDNA synthesis from RNA was performed using μg of RNA, at 50°C The cDNA was then treated with RNase H (Invitrogen) and diluted 1:100 before use for QPCR Quantitative PCR In ... 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 of cells infected ... dendritic cells and alters their immune function J Virol 2003, 77(8):4950-4959 Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real- time quantitative PCR and the 2(-Delta...

Ngày tải lên: 18/06/2014, 18:20

... primed cDNA- synthesis cDNA synthesis from RNA was performed using μg of RNA, at 50°C The cDNA was then treated with RNase H (Invitrogen) and diluted 1:100 before use for QPCR Quantitative PCR In ... 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 of cells infected ... dendritic cells and alters their immune function J Virol 2003, 77(8):4950-4959 Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real- time quantitative PCR and the 2(-Delta...

Ngày tải lên: 20/06/2014, 01:20

... by a dissociation curve and 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) ... apomictic accession, with 98% of aposporous embryo sacs [9] Quantitative real- time PCR (qRT -PCR) has emerged as an important technique to compare the expression profiles of target genes in different ... Selection of reference 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...

Ngày tải lên: 12/08/2014, 03:20

... reaction containing: 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, ... obtained by means of the Cross-Entropy Monte Carlo algorithm present in the software CT values and variability between organs and developmental stages in Mitchell and V30 Real- time PCR reactions ... http://www.biomedcentral.com/1471-2229/10/4 Page of 11 Figure Developmental stages of leaves and flowers used for RNA extractions Representative photographs of leaves and flowers of Petunia hybrida lines Mitchell (a, c) and V30 (b,...

Ngày tải lên: 12/08/2014, 03:21

... 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 were selected, and ... TaqMan real- time PCR for detection of hepatopancreatic parvovirus from Australia J Virol Methods 2007, 140:10-16 42 Hussein IT, Field HJ: Development of a quantitative real- time TaqMan PCR assay ... and application of a one-step real- time Taqman RT -PCR assay for detection of Duck hepatitis virus type1 J Virol Methods 2008, 153:55-60 Deng S, Cheng A, Wang M, Cao P: Serovar-specific real- time...

Ngày tải lên: 12/08/2014, 04:21

... Quantification and validation of 19 genes by qPCR The expression of 19 genes in test lung cancer cell lines (black and gray) and four validation cell lines (white) was quantified by qPCR and compared ... expression of STMN1, AMY2A, and ISL1, and positively correlated with S100P, RAB25, and CDH1 The loading number of component (PC3) was negatively correlated with the expression of RAB25 and AMY2A, and ... each gene and used as the standard sample in the subsequent assay Each standard curve consisted of six serial dilutions (1, 1/5, 1/25, 1/125, 1/625, and 1/3125) of the selected standard cDNA for...

Ngày tải lên: 15/02/2014, 04:20

... design and coordination of the study and helped to draft the manuscript All authors read and approved the final manuscript Material and Methods Virus culture and virus detection by real- time PCR ... primed cDNA- synthesis cDNA synthesis was performed using µg of RNA, at 50°C Finally, cDNA was diluted 1:5 before use in QPCR Quantitative TaqMan PCR Primers, TaqMan probes and QPCR conditions for ... assistance of Delia Barz and Jung-Won Sim-Bandenburg The authors are grateful to Andreas Kurth for critical reading of the manuscript References 10 11 Mackay IM, Arden KE, Nitsche A: Real- time PCR in...

Ngày tải lên: 18/06/2014, 22:20

... design and coordination of the study and helped to draft the manuscript All authors read and approved the final manuscript Material and Methods Virus culture and virus detection by real- time PCR ... primed cDNA- synthesis cDNA synthesis was performed using µg of RNA, at 50°C Finally, cDNA was diluted 1:5 before use in QPCR Quantitative TaqMan PCR Primers, TaqMan probes and QPCR conditions for ... assistance of Delia Barz and Jung-Won Sim-Bandenburg The authors are grateful to Andreas Kurth for critical reading of the manuscript References 10 11 Mackay IM, Arden KE, Nitsche A: Real- time PCR in...

Ngày tải lên: 20/06/2014, 04:20

... 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 ... applicability of real- time PCR for the quantification of GPV because of its remarkable sensitivity and high-throughput potential, which is beyond the scope of other diagnostic methods The real- time PCR ... the fluorescent quantitative real- time PCR (FQ -PCR) , has been used widely to quantify the number of genomic copies of pathogenic microorganisms [18,19] GPV detection by real- time PCR has only been...

Ngày tải lên: 12/08/2014, 04:20

... [21] and macaques [22], all post intramuscular (i.m.) administration of pDNA Gratifyingly, all the studies have given evidence for overall safety as well Quantitative real- time PCR (QRTPCR) is ... authorities will demand a reliable, sensitive and cost effective method for the determination of the amount of residual plasmid and its localization in the body at the time of the slaughter The ... concentration of saline media, size of plasmid DNA, repeated gene transfer However, neither the influence of lag time between plasmid injection and electroporation nor the effect of the age of mice...

Ngày tải lên: 14/08/2014, 19:22

... slope and R value of standard curves for other reactions are listed in Table Specificity of the qRT -PCR qRT -PCR specificities were assessed by melting curve analysis and sequencing of amplified PCR ... Recently, quantitative real time PCR (qRT -PCR) has become widely accepted for rapid and reproducible quantification of gene expression Most previous attempts to quantify Vb expression using qRT -PCR ... and sequencing amplified PCR products Melting curve analysis and sequencing amplified PCR products of reactions for, Cb and Vb subgroups consisting of a single subgroup showed a single peak and...

Ngày tải lên: 18/06/2014, 16:20

... Establishment of the fluorescent quantitative real- time PCR standard curve Establishment of the fluorescent quantitative real- time PCR standard curve Standard curve of the AHV-1 fluorescent quantitative real- time ... of fluorescent quantitative real- time PCR and conventional PCR The real- time PCR was carried out using the ABI AmpliTaq Gold DNA polymerase system with an icycler IQ Realtime PCR Detection System ... 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...

Ngày tải lên: 12/08/2014, 04:21

... Establishment of the fluorescent quantitative real- time PCR standard curve Establishment of the fluorescent quantitative real- time PCR standard curve Standard curve of the AHV-1 fluorescent quantitative real- time ... of fluorescent quantitative real- time PCR and conventional PCR The real- time PCR was carried out using the ABI AmpliTaq Gold DNA polymerase system with an icycler IQ Realtime PCR Detection System ... 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...

Ngày tải lên: 12/08/2014, 04:21

... 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 Patient...

Ngày tải lên: 14/08/2014, 07:21

... for publication of quantitative real- time PCR experiments Clin Chem 55:611–622 Raymaekers M, Smets R, Maes B et al (2009) Checklist for optimization and validation of real- time PCR assays J Clin ... 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) from a research ... strategies commonly 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...

Ngày tải lên: 05/09/2015, 10:35

... for the measurement of the average molecular weight of HMW hyaluronan Analysis of message expression by quantitative realtime RT -PCR Message expression in the synovium of knees and the relative ... levels of the messages (RA) Relative expressionand rheumatoid arthritis for hyaluronanof knees in 2, and -3 and hyaluronidase-1, -2, and -3 in the synovium of knees in osteoarthritis (OA) and rheumatoid ... the production volume of hyaluronan, the elimination volume of hyaluronan from the joint, and the total volume of joint fluid The production of hyaluronan in OA or RA may be decreased because of...

Ngày tải lên: 09/08/2014, 01:24

... Analysis by Real- Time Quantitative PCR Quantitative PCR was performed using the Sequence Detector System, ABI PRISM 7500 (Applied Biosystems 7500 Fast Real- Time PCR System) PCR primers and the TaqMan ... [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 the early detection of tumor ... have been described and can be 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...

Ngày tải lên: 18/06/2014, 16:20

... performance of the PCR was monitored by quantitative real- time PCR (qPCR) The mean cycle threshold (Ct) and the standard deviation of the controls were calculated Samples in which the mean Ct of the ...

Ngày tải lên: 19/06/2014, 08:20

... 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 ... expressed as copies of viral DNA per 100 µL of virus suspension The reliability of real- time PCR was defined by calculating coefficients of variation of Ct values of replicates of standard curve dilutions ... microtiter and transwell plates Semiquantitative polymerase chain reaction (PCR) and quantitative real- time PCR have been recently developed and employed for the detection and quantitation of JCV...

Ngày tải lên: 19/06/2014, 08:20

... performance of the PCR was monitored by quantitative real- time PCR (qPCR) The mean cycle threshold (Ct) and the standard deviation of the controls were calculated Samples in which the mean Ct of the ...

Ngày tải lên: 20/06/2014, 04:20