synthesis rt pcr and quantitative real time pcr

... (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) and pre-designed ... 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 ... real- time RT- PCR Nucleic Acids Res 2001, 29(9):e45 Thellin O, Zorzi W, Lakaye B, De Borman B, Coumans B, Hennen G, Grisar T, Igout A, Heinen E: Housekeeping genes as internal standards: use and...

Ngày tải lên: 18/06/2014, 18:20

... 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 virology ... virus: a comparison of quantitative real- time PCR and plaque assay J Virol Methods 2003, 110:185-191 Nitsche A, Muller CW, Radonic A, Landt O, Ellerbrok H, Pauli G, Siegert W: Human herpesvirus ... HHV-6 experiments and real- time PCR assays and drafted the manuscript ST carried out the CMV experiments HB carried out the YF experiments MM carried out the SARS experiments WS participated in the...

Ngày tải lên: 18/06/2014, 22:20

... (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) and pre-designed ... 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 ... real- time RT- PCR Nucleic Acids Res 2001, 29(9):e45 Thellin O, Zorzi W, Lakaye B, De Borman B, Coumans B, Hennen G, Grisar T, Igout A, Heinen E: Housekeeping genes as internal standards: use and...

Ngày tải lên: 20/06/2014, 01:20

... 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 virology ... virus: a comparison of quantitative real- time PCR and plaque assay J Virol Methods 2003, 110:185-191 Nitsche A, Muller CW, Radonic A, Landt O, Ellerbrok H, Pauli G, Siegert W: Human herpesvirus ... HHV-6 experiments and real- time PCR assays and drafted the manuscript ST carried out the CMV experiments HB carried out the YF experiments MM carried out the SARS experiments WS participated in the...

Ngày tải lên: 20/06/2014, 04:20

... were tested for specificity by RTPCR and also by qRT -PCR, followed by a dissociation curve and agarose gel electrophoresis Real- time PCR conditions and analysis PCR reactions were performed in ... ubiquitin and elongation factor-1 alpha For species with both sexual and apomictic reproductive mode, the best reference genes for qRT -PCR experiments have not been reported yet Real time PCR has ... 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 species, tissues or treatments and also to...

Ngày tải lên: 12/08/2014, 03:20

... 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 a SYBR Green based PCR assay (with ROX as ... results in real- time PCR Nucl Acids Res 2002, 30(9):e36 Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe AFS: Accurate normalization of realtime quantitative RT- PCR data by ... studies by quantitative real- time RT- PCR Molecular Breeding 2009, 23(4):607-616 36 Olsvik PA, Lie KK, Jordal AEO, Nilsen TO, Hordvik I: Evaluation of potential reference genes in real- time RT- PCR studies...

Ngày tải lên: 12/08/2014, 03:21

... fluorescent quantitative real- time PCR (FQ -PCR) This technology accurately quantifies target DNA in a given sample and then could accurately detect viral loads in clinical samples[34] Yang and Guo ... proviral load by a TaqMan real- time PCR assay J Virol Methods 2002, 102:37-51 38 Yun Z, Lewensohn-Fuchs I, Ljungman P, Ringholm L, Jonsson J, Albert J: A real- time TaqMan PCR for routine quantitation ... 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...

Ngày tải lên: 12/08/2014, 04:21

... cancer 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 ... data, and manuscript preparation TM: Experiments (cell culture and DNA microarray analysis) YD: Experiments (cell culture and quantitative real- time PCR analysis) YF: Experiments (cell culture and ... tissues and quantitative RT- PCR reliability Exp Mol Pathol 2009, 87:146-51 Provenzano M, Mocellin S: Complementary techniques: validation of gene expression data by quantitative real time PCR Adv...

Ngày tải lên: 15/02/2014, 04:20

... 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 ... 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 ... the basic assay and performed most experiments including cloning, protein purification, cell preparation and stimulation, qRT -PCR, and data analysis JYP helped to perform qRT -PCR and interpret...

Ngày tải lên: 18/06/2014, 16:20

Ngày tải lên: 14/08/2014, 07:21

... studies have given evidence for overall safety as well Quantitative real- time PCR (QRTPCR) is the most widely used method for specific quantitative assay of ultra low concentration of pDNA in ... http://www.gvt-journal.com/content/6/1/11 Conclusion Quantitative real- time PCR (QRTPCR) assay was developed to assess a residual pDNA vaccine pVAX-Hsp60 TM814 in mice and beef cattle In beef cattle, ultra ... QRTPCR, participated in quantification of pDNA, and participated in preparation of the manuscript VK participated in preparation of cationic liposomes, carried out the histology experiments and...

Ngày tải lên: 14/08/2014, 19:22

... of qPCR: A Mature Technology? Alessandro Raso and Roberto Biassoni Abstract Quantitative PCR is the “gold standard” technology to quantify nucleic acids and, since the first report describing real- time ... University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK For further volumes: http://www.springer.com/series/7651 Quantitative Real- Time PCR Methods and Protocols Edited by Roberto Biassoni ... characterized by slowdown trend of amplification and the products Roberto Biassoni and Alessandro Raso (eds.), Quantitative Real- Time PCR: Methods and Protocols, Methods in Molecular Biology, vol...

Ngày tải lên: 05/09/2015, 10:35

... Heid CA, Stevens J, Williams PM: Real time quantitative PCR Genome Res 1996, 6:986-994 36 Gibson UEM, Heid CA, Williams A: Novel method for real time quantitative RT- PCR Genome Res 1996, 6:995-1001 ... RA were determined by realtime RT- PCR in accordance with the manufacturer's instructions and reported methods [35-39] The gene-specific PCR oligonucleotide primer pairs and gene-specific oligonucleotide ... osteoarthritis hyaluronidase-1, -2, and -3 in the synovium synthase-1, 2, and -3 and (OA) levels of the messages (RA) Relative expressionand rheumatoid arthritis for hyaluronanof knees in 2, and...

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 ... carried out the real- time RT- PCR, participated in the clinical data collecting of the gastric carcinoma patients and drafted the manuscript LZH carried out the real- time RT- PCR ZZW participated in ... tumor cells by real- time RT- PCR of mRNA markers It shows the positive rate of mRNA markers from literature for detection of tumor cells by real- time RT- PCR Abbreviations RT- PCR: Reverse Transcriptase-Polymerase...

Ngày tải lên: 18/06/2014, 16:20

... 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 DNA ranging ... GPV detection by real- time PCR has only been reported by Bi [20]; in that study, the method was not optimized and a FQ -PCR standard curve was not generated In this study, we reported the optimization ... factors could not be assessed by qualitative PCR [16,17] Establishment of FQ -PCR standard curve The FQ -PCR amplification curves and the corresponding FQ -PCR standard curve (Figure 1) were generated...

Ngày tải lên: 12/08/2014, 04:20

... doi:10.1186/1471-2229-10-49 Cite this article as: Artico et al.: Identification and evaluation of new reference genes in Gossypium hirsutum for accurate normalization of real- time quantitative RT- PCR data BMC Plant ... gene expression and internal standard selection Biotechnology Advances 2009, 27(4):323-333 12 Andersen CLJJ, Orntoft TF: Normalization of real- time quantitative reverse transcription -PCR data: A ... quantification realtime PCR Bmc Plant Biol 2008, 9:59 44 Frost PNF: Validation of reference genes for transcription profiling in the salmon louse, Lepeophtheirus salmonis, by quantitative real- time PCR...

Ngày tải lên: 12/08/2014, 03:21

... Paisley, UK) and incubated at 37°C for hour Two μl of cDNA were used for each PCR amplification by "real time" technology (7300 Real Time PCR system, Applied Biosystems, Rotkreuz, Switzerland) according ... http://www.translational-medicine.com/content/6/1/58 Authors' contributions ES-T and NR performed qRT -PCR assays in WB, DMF designed the study and participated in the performance of qRT -PCR assays and in writing the paper DM helped collecting ... vaccinated donors and performed serological studies CF-M helped designing the qRT -PCR strategy GCS provided funding and helped designing the study and writing the paper PZ designed the qRT -PCR strategy,...

Ngày tải lên: 18/06/2014, 15:20

... 2008, 6:61 Part 2: IFNγ real time RT- PCR validation Specificity IFNγ real time RT- PCR specificity is defined as lack of response to irrelevant peptides and HIV negative control peptide and positive ... to TIL/1000 PBMC) (B) Expanded dose response: Graph in (A) is expanded to show details at the lower doses Figure IFNγ real time RT- PCR specificity IFNγ real time RT- PCR specificity TIL cells ... http://www.translational-medicine.com/content/6/1/61 Figure IFNγ real time RT- PCR standard curve (linearity) IFNγ real time RT- PCR standard curve (linearity) Linear response (IFNγ plasmid copy number...

Ngày tải lên: 18/06/2014, 15:20

... with automation to further improve the timeframe for genotyping a NoV infection Conclusion A real time RT- PCR assay and a RLBH assay were developed and utilised to identify and genotype the causative ... with three previously published RT- PCR assays [7-9] A Real- Time SYBR green RT- PCR assay was then developed based on the primer pairs to detect and quantify both GI and GII NoV in the Irish population ... of human NoV by Real- Time RT- PCR 56 samples were taken from two outbreaks of NoV in two Irish hospitals in 2005 and 2006 These samples were applied to the GII NoV real- time assay and 26 samples...

Ngày tải lên: 18/06/2014, 18:20

... with automation to further improve the timeframe for genotyping a NoV infection Conclusion A real time RT- PCR assay and a RLBH assay were developed and utilised to identify and genotype the causative ... with three previously published RT- PCR assays [7-9] A Real- Time SYBR green RT- PCR assay was then developed based on the primer pairs to detect and quantify both GI and GII NoV in the Irish population ... of human NoV by Real- Time RT- PCR 56 samples were taken from two outbreaks of NoV in two Irish hospitals in 2005 and 2006 These samples were applied to the GII NoV real- time assay and 26 samples...

Ngày tải lên: 20/06/2014, 01:20