... isoforms of HAS and hyaluronidase in knee synovium obtained from control donors and patients with OA or RA, by quantitative reversetranscriptasepolymerasechainreaction (RT -PCR) , in order to ... Society [32] SD, standard deviation Table Sequences of the gene-specific oligonucleotide primers and probes for real-time reversetranscriptasepolymerasechainreaction Primer and probe HAS-1 Sequence ... -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 -3 and hyaluronidase-1, -2, and...
... developed an RT -PCR assay and Taqman RT -PCR assay for WEEV and EEEV [11] But none of the above methods was available for detecting and distinguishing WEEV and EEEV in a single reaction In this ... containment Reverse- transcriptasePCR (RT -PCR) is a rapid and sensitive method that is being increasingly used as an adjunct to serology for arbovirus diagnosis [11,3] Real-time RT -PCR is more ... uL PCR was performed with a LightCycler 2.0 (Roche, Switzerland) The reactions were incubated at 50°C for 30 min, followed by 95°C for 10 (inactivation reverse transcriptase/ activation Taq polymerase) ,...
... 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- PolymeraseChain Reaction; ... colonic cells in peripheral blood of colorectal cancer patients by means of reversetranscriptaseandpolymerasechainreaction Br J Cancer 1998, 78:1368-1372 31 Schlimok G, Funke I, Pantel K, ... T: Prediction of peritoneal micrometastasis by peritoneal lavaged cytology andreverse transcriptase- polymerasechainreaction for matrix metalloproteinase-7 mRNA Clin Cancer Res 2001, 7:1647-1653...
... HD, Grant PR, Garson JA, Tedder RS, Chandy GM, Abraham P: Quantitation of hepatitis C virus using an in-house real-time reversetranscriptasepolymerasechainreaction in plasma samples Diagn Microbiol ... Huang J, Yang CM, Wang LN, Meng S, Deng W, Li JM: A novel real-time multiplex reverse transcriptase- polymerasechainreaction for the detection of HIV-1 RNA by using dual-specific armored RNA as ... 10.1186/1743-422X-7-117 Cite this article as: Meng and Li, A novel duplex real-time reverse transcriptase- polymerasechainreaction assay for the detection of hepatitis C viral RNA with armored RNA as...
... being dengue positive and selected for confirmation and serotyping by reversetranscriptasepolymerasechainreaction In 2003, 45 specimens were analysed of which 14 were males and 31 were females ... virus However, single-step reversetranscriptasepolymerasechainreaction (RT -PCR) detection and typing of dengue virus offers a sensitive, specific, reproducible and rapid alternative that ... minikit Reverse transcription andpolymerasechainreaction was performed using the One-Step Superscript III/RT/Platinum Taq Mix (Invitrogen), 0.1 M dithiothreitol (DTT), 5' primer D1 and 3' primer...
... Methods Quantitative Reverse Transcriptase- PolymeraseChainReaction (RT -PCR) Real-time quantitative RT -PCR analysis of gene expression [13,14] was carried out on first-strand cDNA derived from ... Osada T, Uehara H, Kim H and Ikai A mRNA analysis of single living cells J Nanobiotechnology 2003, 1:2 Grillo M and Margolis FL Use of reversetranscriptasepolymerasechainreaction to monitor expression ... R, Dinnebier G and Jaross W In situ reverse transcriptase- nested polymerasechainreaction to identify intracellular nucleic acids without the necessity of DNAse pretreatment and hybridisation...
... 55/lizarb/oriezurc/42A rof ecneuqes ehT sniarts 21 eht rof dezylana saw noiger tegrat ehT )ASU ,ratsanD( erawtfos ratsanD eht gnisu dengila dna )1 elbaT( A C aisA 2TAS O O O O O O O O 55/lizarB/oriezurC/42A ... eht gnitarapes ybereht ,esaremylop AND qaT fo ytivitca esaelcunoxe eht ot eud dedarged si eborp eht ,srucco noitacifilpma sA tegrat RCP eht nihtiw ecneuqes AND dnarts-elgnis yratnemelpmoc sti...
... in a 25 µl PCR The remainder of the reactions were made up by µl PCR Buffer (Gibco BRL, Grand Island, NY), 0.25 µg of each primer, mM MgC12 in each CD4 reactionand mM MgC12 for CD8 reactions, ... Quantitative methods for reversetranscriptasepolymerasechainreaction (RT -PCR) are rapidly surpassing all other methods of quantifying mRNA levels We therefore set out to develop an RT -PCR based method ... correlation between CD4 and CD8 cell counts and mRNA levels of CD4 and CD8 could be determined by QC-RT -PCR There was also no correlation between CD4 and CD8 mRNA levels and the number of cells...
... (H5N1) viruses and detection of anti-H5 antibody among household and social contacts”, J Infect Dis, 180, pp 1763-70 36 Lamb RA and Krug RM (2001), “Orthomyxoviridae: The Viruses and Their Replication”, ... fixation and hemagglutination inhibition tests in the diagnosis of influenza A and B virus infections”, Purified hemagglutinin in subtype-specific diagnosis, J Vir.Methods 10, 75-84 34 Kandun IN, ... của , virus cúm A/H5N1 thực hiê ̣n cùngmột phản ứng RT -PCR Phản ứng m utiplex RT -PCR: Được thực với sinh phẩm Qiagen OneStep RT -PCR theo hướng dẫn của nhà sả n xuấ t máy luân nhiê...
... deactivate reverse transcriptase; 45 or 50 cycles of 10 s at 95oC to denature and 20 s at 55oC to anneal and 20 s at 72oC for extension and at 72oC for final extension Reverse transcription andPCR ... TCID50 /reaction to 2.24 × 101.0 TCID50 /reaction Real-time RT -PCR assay could detect JE RNA ranging from 1.12 × 106.0 to 1.12 × 101.0 TCID50/ml Therefore, detection limits of RT -PCR and real-time RTPCR ... isolation and real-time RT -PCR -/-: Negative results in both virus isolation and real-time RT -PCR 350 Dong Kun Yang et al has several advantages over conventional PCR First, realtime RT -PCR yields...
... immediately for pp65, and real time PCR for HCMV (qRT- PCR) The study was approved by both the institute’s research and ethics committee and informed consent was obtained from the donors and the renal ... Madhavan et al.: “pp65 antigenemia and real time polymerasechainreaction (PCR) based-study to determine the prevalence of human cytomegalovirus (HCMV) in kidney donors and recipients with follow-up ... antigenemia and real - time PCR The highest copy number detected by real - time PCR was 29, 768 copies/ml and lowest was copies/ml The highest positive pp65 cell count recorded among them was 78 cells and...
... học phân tử 20 1.3.3.1.Phương pháp RT – PCR (Reverse – transcription polymerasechain reaction) 20 1.3.3.3 Các thành phần phản ứng PCR 23 CHƢƠNG 2: VẬT LIỆU VÀ PHƢƠNG PHÁP NGHIÊN ... – PCR (Reverse – transcription polymerasechain reaction) Phương pháp tạo RT- PCR gồm bước chính: - Tách chiết RNA từ bệnh phẩm biểu mô - Tạo cDNA phản ứng chép ngược - Thực phản ứng PCR với ... triphosphate dCTP: Deoxycytidine triphosphate UTRs: Untranslated regions RT -PCR: Reverse transcription polymerasechainreaction kDa: kilo dalton H&E: Hematoxylin EoSin LỜI MỞ ĐẦU Ngày việc khai...
... mẫu PolymeraseChainReaction Phản ứng chuỗi trùng hợp Reverse Transcription PolymeraseChainReaction Phản ứng chuỗi đồng phân hóa chép ngƣợc mRTPCR Multiplex Reverse Transcription PolymeraseChain ... máu bệnh phẩm có hiệu suất tốt với tỷ lệ phát cao [39] Phương pháp RT -PCR RT -PCR (reverse transcriptasepolymerasechain reaction) phƣơng pháp dùng để khuếch đại cDNA đƣợc tạo từ RNA dựa vào ... pháp single PCR (sPCR) 20-200 pcg mPCR Thí nghiệm đƣợc thực nồng độ pha loãng 10x mẫu có chứa tất virus để xác định giới hạn phát multiplex RT -PCR Độ nhậy phƣơng pháp mPCR so với sPCR tƣơng ứng...
... Omniscript and Sensiscrip Reverse Transcriptases tiếp đến phản ứng PCR nhờ enzyme Hot Star Taq DNA polymerase Như vậy, tất thành phần phản ứng RT PCR cho vào tube phản ứng lần Thể tích phản ứng RT -PCR ... 0.6 µM 0.7 µM band sản phẩm RT -PCR rõ nét xuất băng phụ dimer rõ phía Ở nồng độ mồi 0.5 µM phản ứng cho band sáng rõ nét band phụ dimer mờ Hình 3.5B: Phản ứng RT -PCR phát gen HA, band sản phẩm ... µM band sáng rõ band phụ dimer xuất nhiều so với nồng độ 0.4 µM Còn nồng độ mồi khác sản phẩm RT -PCR thấp (các band mờ hơn) Hình 3.5C: Phản ứng RT -PCR phát gen NA nồng độ mồi 0.5 µM cho band...
... Total RNA (1 μg) and random 8-mer primers (50 μM) were denatured at 65°C for and cooled down on ice To synthesise the first-strand cDNA, the RNA and primers were mixed in × reaction buffers, ... 21:844-849 21 Si W, Zhou S, Wang Z, Cui S: A multiplex reverse transcription-nested polymerasechainreaction for detection and differentiation of wildtype and vaccine strains of canine distemper virus ... the first round amplification was conducted by polymerasechainreaction (PCR) with the outer primers CDF-F: 5'-AGAGTGCAAAATAGTAAGAATCCAAGC-3' and CDF-R: 5'-GAAAGAGACTGGCTATTCCGATGC-3', which...
... 4°C Amplified Methylation Polymorphism PolymeraseChainReaction (AMP PCR) The PCRreaction consisted of DNA sample (genomic or digested template), Taq polymerase enzyme (AmpliTaq® Stoffel fragment, ... clear and reproducible marker bands were counted, and the comparison of bands between genomic and digested templates was done according to the appearance of the marker mentioned above Number and ... the PCR to increase the sensitivity of marker detection Conclusions We applied an arbitrarily primed PCR- based technique, Amplified Methylation Polymorphism PolymeraseChainReaction (AMP PCR) ,...
... synthesis reaction was performed by polymerasechainreaction as described elsewhere using primer Moloney murine leukemia virus reversetranscriptase (M-MLV RT)[6-10] The first-round PCR was performed ... RT -PCR and nested PCR in the diagnosis of canine distemper Vet Res Commun 2005, 29:347-59 10 Li JZ, He HB, Xia XZ, et al.: Establishment and application of reverse transcription Polymerasechain ... chosen for the RT-nPCR Specificity of the multiplex RT-nPCR A fragment of 247 and 177 bp was amplified from CDV wild-type strain and the vaccine strain, respectively Two bands of 247 and 177 bp were...
... typically performed in a 50µl reaction mix containing 0.25nmoles of forward andreverse primers, 2mM final concentration of dNTP, 1.5 U of Taq polymerase in 1X buffer and proteinase K digested sample ... scrofulaceum and M intracellulare PCRand microscopy were negative for both samples identified as MOTT and positive for 98% (47/48) of the remaining samples (Table 1A, serial-a) Table 1A: Efficiency of PCR ... pneumonia and cancer lung, one each of post CABG, COPD and lung abscess and 12 with cough and cold All samples from these individuals had tested negative for tuberculosis by all three diagnostics No PCR...
... endonuclease cleavage sites Polymerase chain reaction, cycle Primer Primer Template Cycle 1 Denature Anneal primers Synthesize new DNA with polymerasePolymerasechain reaction, cycle Cycle Denature ... produced on sucrose media: positive selection for recombinants PCR provides access to specific DNA segments • PolymeraseChainReaction • Requires knowledge of the DNA sequence in the region ... new DNA with polymerase PCR, cycle Cycle (focus on DNA segments bounded by primers) Denature Anneal primers Synthesize new DNA with polymerase duplex molecules of desired product PCR, cycle 4:...
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