qpcr and reverse transcriptase polymerase chain reaction qrt pcr

... isoforms of HAS and hyaluronidase in knee synovium obtained from control donors and patients with OA or RA, by quantitative reverse transcriptase polymerase chain reaction (RT -PCR) , in order to ... Society [32] SD, standard deviation Table Sequences of the gene-specific oligonucleotide primers and probes for real-time reverse transcriptase polymerase chain reaction 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...

Ngày tải lên: 09/08/2014, 01:24

... 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- transcriptase PCR (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) ,...

Ngày tải lên: 12/08/2014, 01:22

... 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 Chain Reaction; ... colonic cells in peripheral blood of colorectal cancer patients by means of reverse transcriptase and polymerase chain reaction Br J Cancer 1998, 78:1368-1372 31 Schlimok G, Funke I, Pantel K, ... T: Prediction of peritoneal micrometastasis by peritoneal lavaged cytology and reverse transcriptase- polymerase chain reaction for matrix metalloproteinase-7 mRNA Clin Cancer Res 2001, 7:1647-1653...

Ngày tải lên: 18/06/2014, 16:20

... HD, Grant PR, Garson JA, Tedder RS, Chandy GM, Abraham P: Quantitation of hepatitis C virus using an in-house real-time reverse transcriptase polymerase chain reaction in plasma samples Diagn Microbiol ... Huang J, Yang CM, Wang LN, Meng S, Deng W, Li JM: A novel real-time multiplex reverse transcriptase- polymerase chain reaction 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- polymerase chain reaction assay for the detection of hepatitis C viral RNA with armored RNA as...

Ngày tải lên: 12/08/2014, 04:20

... being dengue positive and selected for confirmation and serotyping by reverse transcriptase polymerase chain reaction In 2003, 45 specimens were analysed of which 14 were males and 31 were females ... virus However, single-step reverse transcriptase polymerase chain reaction (RT -PCR) detection and typing of dengue virus offers a sensitive, specific, reproducible and rapid alternative that ... minikit Reverse transcription and polymerase chain reaction was performed using the One-Step Superscript III/RT/Platinum Taq Mix (Invitrogen), 0.1 M dithiothreitol (DTT), 5' primer D1 and 3' primer...

Ngày tải lên: 12/08/2014, 04:21

... Methods Quantitative Reverse Transcriptase- Polymerase Chain Reaction (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 reverse transcriptase polymerase chain reaction to monitor expression ... R, Dinnebier G and Jaross W In situ reverse transcriptase- nested polymerase chain reaction to identify intracellular nucleic acids without the necessity of DNAse pretreatment and hybridisation...

Ngày tải lên: 11/08/2014, 00:21

... 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...

Ngày tải lên: 07/08/2014, 18:21

... 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 reaction and mM MgC12 for CD8 reactions, ... Quantitative methods for reverse transcriptase polymerase chain reaction (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...

Ngày tải lên: 11/08/2014, 08:20

... (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...

Ngày tải lên: 10/02/2014, 20:39

... 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 and PCR ... 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...

Ngày tải lên: 07/08/2014, 18:20

... 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 polymerase chain reaction (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...

Ngày tải lên: 12/08/2014, 02:20

... học phân tử 20 1.3.3.1.Phương pháp RT – PCR (Reverse – transcription polymerase chain reaction) 20 1.3.3.3 Các thành phần phản ứng PCR 23 CHƢƠNG 2: VẬT LIỆU VÀ PHƢƠNG PHÁP NGHIÊN ... – PCR (Reverse – transcription polymerase chain 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 polymerase chain reaction kDa: kilo dalton H&E: Hematoxylin EoSin LỜI MỞ ĐẦU Ngày việc khai...

Ngày tải lên: 31/08/2014, 07:43

... mẫu Polymerase Chain Reaction Phản ứng chuỗi trùng hợp Reverse Transcription Polymerase Chain Reaction Phản ứng chuỗi đồng phân hóa chép ngƣợc mRTPCR Multiplex Reverse Transcription Polymerase Chain ... máu bệnh phẩm có hiệu suất tốt với tỷ lệ phát cao [39] Phương pháp RT -PCR RT -PCR (reverse transcriptase polymerase chain 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...

Ngày tải lên: 30/11/2015, 20:08

... 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...

Ngày tải lên: 17/06/2016, 22:59

... 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 polymerase chain reaction for detection and differentiation of wildtype and vaccine strains of canine distemper virus ... the first round amplification was conducted by polymerase chain reaction (PCR) with the outer primers CDF-F: 5'-AGAGTGCAAAATAGTAAGAATCCAAGC-3' and CDF-R: 5'-GAAAGAGACTGGCTATTCCGATGC-3', which...

Ngày tải lên: 12/08/2014, 04:20

... 4°C Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) The PCR reaction 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 Polymerase Chain Reaction (AMP PCR) ,...

Ngày tải lên: 12/08/2014, 18:22

... synthesis reaction was performed by polymerase chain reaction as described elsewhere using primer Moloney murine leukemia virus reverse transcriptase (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 Polymerase chain ... 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...

Ngày tải lên: 12/08/2014, 04:20

... typically performed in a 50µl reaction mix containing 0.25nmoles of forward and reverse primers, 2mM final concentration of dNTP, 1.5 U of Taq polymerase in 1X buffer and proteinase K digested sample ... scrofulaceum and M intracellulare PCR and 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...

Ngày tải lên: 06/03/2014, 04:20

... endonuclease cleavage sites Polymerase chain reaction, cycle Primer Primer Template Cycle 1 Denature Anneal primers Synthesize new DNA with polymerase Polymerase chain reaction, cycle Cycle Denature ... produced on sucrose media: positive selection for recombinants PCR provides access to specific DNA segments • Polymerase Chain Reaction • 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:...

Ngày tải lên: 13/03/2014, 18:39

... yticificepS giF AND fo gp 05 ;5 enaL ,AND fo gp 001 ;4 enaL ,AND fo gn ;3 enaL ,AND fo gn 01 ;2 enaL ,AND etalpmet fo gn 001 ;1 enaL ,III eaH yb detsegid 471Xø ;M detulid yllaires saw AND etalpmet ... stnemgarf AND deifilpma cificepsnon yna ecudorp ton did dna ,sniarts lairetcab hcae deifirup AND etalpmet htiw tnemgarf AND detcepxe eht decudorp yassa RCP ehT snoitcaer RCP hcae ni AND etalpmet ... dna muirumihpyt S rof AND deifirup fo gp 001 sa elttil sa tceted dluoc yassa RCP elgnis ehT gp 01 ot gn 001 morf AND deifirup hcae fo snoitulid laires no detcudnoc saw RCP ,AND etalpmet hcae fo...

Ngày tải lên: 07/08/2014, 18:21