ifng real time rt pcr validation

... citation purposes) Journal of Translational Medicine 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 ... 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 at different numbers were spiked into 106 PBMC; ... 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...

Ngày tải lên: 18/06/2014, 15:20

... of real- time RT- PCR Since the advent of real- time RT- PCR the number of applications has grown exponentially It is outside the scope of this book to describe every application of real- time RT- PCR; ... mRNA targets of Real- time RT- PCR 223 Table 9.2 Real- time thermal cyclers PCR system Company Excitation specifications Detection specification Applied Biosystems 7300 Real- Time PCR System Applied ... are a range of real- time PCR instruments and the properties of some of these are summarized in Table 9.2 The quantification of results from a real- time RT- PCR experiment is important and there...

Ngày tải lên: 25/10/2013, 22:20

... đường chuẩn cho phương pháp định lượng real time PCR chẩn đoán PRRSV Đoạn mồi phản ứng realtime PCR sử dụng EvaGreen Mồi F7 R7 cho phản ứng realtime PCR theo Martınez cs, 2007 Trình tự mồi vị trí ... lượng PRRSV kỹ thuật realtime sử dụng mẫu dò (probe) Trong đó, kỹ thuật realtime PCR sử dụng thuốc nhuộm kỹ thuật định lượng tránh vấn đề Do đó, nghiên cứu quy trình realtime RT- PCR với EvaGreen ... hiệu phản ứng real time RT- PCR định lƣợng PRRSV Để kiểm tra tính đặc hiệu phản ứng, thực phản ứng real time RT- PCR với mẫu dương tính PCV âm tính với PRRSV (được xác định phương pháp PCR) , mẫu dương...

Ngày tải lên: 25/03/2014, 04:21

... quantification 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 ... TCG CT CCT CCC TTT GGA ACC AGT TGC shown) 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 ... outbreak (Fig 4A and 4B) detected by Real- time RT- PCR revealed that all 26 of the samples were genotype GII/4 This result was confirmed by sequencing of the RT- PCR products The predominance of the...

Ngày tải lên: 18/06/2014, 18:20

... quantification 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 ... TCG CT CCT CCC TTT GGA ACC AGT TGC shown) 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 ... outbreak (Fig 4A and 4B) detected by Real- time RT- PCR revealed that all 26 of the samples were genotype GII/4 This result was confirmed by sequencing of the RT- PCR products The predominance of the...

Ngày tải lên: 20/06/2014, 01:20

... fo stimil eht rof hcraes ruo ni lortnoc evitisop a sa desu saw gurd hcae fo noitulos lm/gµ 05 a dna lortnoc evitagen eht sa desu saw retaw dellitsiD selpmas lortnoC la te idnatihS olakanA 871 ... ,lm/gµ naht retaerg snoitartnecnoc ta tceted ylno dluoc ti hcihw ,revewoh ,muilorpma ot ytivitisnes fo kcal a detartsnomed msinagro siht ,yduts ruo nI lm/gµ naht rewllams snoitartnecnoc ta enodilozaruf ... deviecer noitartnecnoc gurd hcae fo etacilper a gniniatnoc ebut hcae taht hcus sebut eht ot dedda saw tnatanrepus ehT noitulos fo tuo llaf ot sirbed eussit wolla ot nim rof degufirtnec erew selpmas...

Ngày tải lên: 07/08/2014, 18:21

... channel Real time RT- PCR optimization The concentration of primers and probes was optimized in separate PCRs (two primers, one probe) and re-adjusted when combined in the multiplex RT- qPCR to ... samples pretested by M-specific RT- qPCR [14] to be positive for AIV was examined using the H5 multiplex RT- qPCR assay and the generic H5 protocol [23] The multiplex RT- qPCR found all 50 samples positive ... birds in Egypt in 2006- 2010 by multiplex H5 RT- qPCR and a generic H5specific RT- qPCR [20] Additional file 2: Analytical specificity of the multiplex RT- qPCR for Egyptian HPAIV H5N1 using different...

Ngày tải lên: 12/08/2014, 01:22

... control (Fig and Fig 3) Comparison of real- time qRT -PCR and conventional RT- PCR Next, the detection limit of the real- time qRT -PCR assay and the conventional RT- PCR assay were compared The Table ... Diagnostic field samples tested positive by real- time qRT -PCR or conventional RT- PCR Tissue samples Number Conventional RT- PCR positive Real- time qRT -PCR positive Lung 40 36 39 Lymphoid tissues ... 72°C segment in the PCR step The results were analysed by using MxPro™ QPCR Software One step SYBR green real- time RT- PCR Melting curve analysis One step SYBR green real- time RT- PCR amplification...

Ngày tải lên: 12/08/2014, 04:20

... tests All experiments were done in duplicate Real- time RT- PCR Real- time RT- PCR was performed using iScript™ OneStep RT- PCR Kit Probes in Chromo real time PCR machines (Bio-Rad, Hercules, CA, USA) ... Page of conventional RT- PCR, rRT -PCR has a smaller risk of cross-contamination, higher sensitivity and specificity, and shorter per sample laboratory turnaround time Several rRT -PCR assays for H5N1 ... influenza virus by TaqMan-MGB real- time RT- PCR Lett Appl Microbiol 2008, 46(1):20-25 19 Chen W, He B, Li C, Zhang X, Wu W, Yin X, Fan B, Fan X, Wang J: Real- time RT- PCR for H5N1 avian influenza...

Ngày tải lên: 12/08/2014, 04:21

... sensitivity and reproducibility between different compared RT- PCR settings Therefore the control can be used to optimize a complete real- time RT- PCR procedure from extraction to analysis during method ... EasyMAG extractor and real- time RT- PCR analysis on LightCycler 480 showed a higher sensitivity and precision and yielded a lower limit of detection as compared to our earlier reported assay In addition, ... of Clinical Microbiology 2001, 39:4093-4096 Verstrepen WA, Bruynseels P, Mertens AH: Evaluation of a rapid real- time RT- PCR assay for the detection of enterovirus RNA in cerebrospinal fluid specimens...

Ngày tải lên: 12/08/2014, 04:21

... sensitivity and reproducibility between different compared RT- PCR settings Therefore the control can be used to optimize a complete real- time RT- PCR procedure from extraction to analysis during method ... EasyMAG extractor and real- time RT- PCR analysis on LightCycler 480 showed a higher sensitivity and precision and yielded a lower limit of detection as compared to our earlier reported assay In addition, ... of Clinical Microbiology 2001, 39:4093-4096 Verstrepen WA, Bruynseels P, Mertens AH: Evaluation of a rapid real- time RT- PCR assay for the detection of enterovirus RNA in cerebrospinal fluid specimens...

Ngày tải lên: 12/08/2014, 04:22

... number of real- time RT- PCR experiments focusing on grape berries has been published Based on microarray and real- time RT- PCR data, UBQ-L40 [14] and one paralog of EF1-α [2] were previously reported ... inappropriate reference gene for real- time reverse transcription PCR data normalization Analytical Biochemistry 2005, 344:141-143 Huggett J, Dheda K, Bustin S, Zumla A: Real- time RT- PCR normalisation; strategies ... quantitative real- time polymerase chain reaction (PCR) data Neuroscience Letters 2003, 339:62-66 Pfaffl MW: A new mathematical model for relative quantification in real- time RT- PCR Nucleic Acids...

Ngày tải lên: 12/08/2014, 05:20

... biển gene cytokine: - Phương pháp RT- PCR real- time RT- PCR: phương pháp phản ánh mức độ biểu gene mã hóa cho cytokine mức phiên mã Những năm gần real- time RT- PCR ngày sử dụng nhiều để định lượng ... cs, 2007) Trong đề tài này, sử dụng kỹ thuật real- time RT- PCR để phát thay đổi mức độ biểu gene cytokine quan trọng hoạt động miễn dịch Vì real- time RT- PCR sử dụng rộng rãi để định lượng cytokine ... GraphPad Prism 29 3.2 Áp dụng quy trình real- time RT- PCR với số chất thử nghiệm Đầu tiên, kiểm tra khả phát biểu gene IL-2 TNF-α quy trình real- time RT- PCR vừa xây dựng với hai chất làm chứng...

Ngày tải lên: 07/02/2015, 15:05

... nhân gây BTCM EV71 kỹ thuật Realtime RT- PCR Định chuẩn xét nghiệm Real- time RT- PCR Qui trình chẩn đoán EV71 ba bước gồm phản ứng RT- PCR kết hợp với phản ứng Real- time PCR nghiên cứu chứng minh ... Phản ứng Real- time RT- PCR xem xác có tương đồng với kết phản ứng RT- PCR định tính EV71 Phản ứng Real- time RT- PCR sử dụng đoạn mồi EV có độ xác tốt bắt buột phải dương tính phản ứng RT- PCR mẫu ... phát triển kỹ thuật Real- time RT- PCR rút ngắn thời gian trả kết nhanh so với RT- PCR cổ điển (2 so với 10 giờ) Các nghiên cứu Đài loan Singapore [12], [17] cho thấy Real- time RT- PCR có độ nhạy cảm...

Ngày tải lên: 07/02/2015, 23:32

... 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 to ... 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 ... antigen specific cytotoxicity and lymphoproliferation assays or quantitative real- time polymerase chain reaction (qRT -PCR) for the detection of cytokine gene expression [1-3] These methods generally...

Ngày tải lên: 18/06/2014, 15:20

... data using real- time quantitative PCR and the 2(-Delta Delta C(T)) method Methods 2001, 25:402-408 Pfaffl MW: A new mathematical model for relative quantification in real- time RT- PCR Nucleic ... Stevens J, Livak KJ, Williams PM: Real time quantitative PCR Genome Res 1996, 6:986-994 Nolan T, Hands RE, Bustin SA: Quantification of mRNA using real- time RT- PCR Nat Protocols 2006, 1:1559-1582 ... for the exchange of qPCR data under the form of XML files [15] Data can be imported from a number of data formats Two standards (qBase internal format and RDML (Real- time PCR Data Markup Language))...

Ngày tải lên: 14/08/2014, 17:22

... mẫu sản phẩm real- time PCR cho kết real- time PCR [+] làm tinh để thu nhận sản phẩm PCR với thử nghiệm tinh sản phẩm PCR (PCR clean-up kit) Promega sản xuất (Cat #A9281) Sản phẩm PCR sau tinh ... minh họa kết điện di sản phẩm PCR sau tinh rõ nét, dù nồng độ HCV số mẫu thấp Điều chứng tỏ qui trình RT real- time PCR mà xây dựng tảng dùng thử nghiệm RT TQ real- time PCR phát định lượng HCV-RNA ... genotype HCV giải trình tự trực tiếp sản phẩm PCR phản ứng real- time PCR dùng taqman probe mà xây dựng đạt độ nhạy cao tương đương phương pháp RT TQ real- time PCR mà công ty Nam Khoa phát triển sử dụng...

Ngày tải lên: 01/08/2014, 03:21

... 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 ... 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 ... 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...

Ngày tải lên: 12/08/2014, 03:21

... Accurate normalization of real- time quantitative RT- PCR data by geometric averaging of multiple internal control genes Genome Biol 2002, 3:7 Gachon CMA, Charrier B: Real- time PCR: what relevance to ... salmonis, by quantitative real- time PCR Vet Parasitol 2003, 118:169-174 45 Robinson TLSI, Sutherland J: Validation of candidate bovine reference genes for use with real- time PCR Vet Immunol and Immunopathol ... by real- time PCR Bmc Plant Biology 2008, 8:112 Jain MNA, Tyagi AK, Khurana JP: Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real- time...

Ngày tải lên: 12/08/2014, 03:21