rt pcr in cdna cloning

... 062 ta tinu ecnabrosba htiw sa noitnevnoc eht gnisu sisylana cirtemotohp -ortceps yb deifitnauq saw ANR locotorp desab tnegaer IRT gnisu detcartxe saw selpmas eussit morf ANR latoT noitcartxe ANR ... dezisehtnys erew sANDc dnarts tsriF RCP-TR reffub gninnur leg sa SPOM %1 gnisu siserohportcele leg esoraga edyhedlamrof %1 yb dessessa saw ANR detcartxe fo ytilauq ehT ytirup dna noitartnecnoc elpmas fo ... 14 63 13 62 14 63 13 62 elcyC lortnoC tnemtaerT eneG deerB )lµ 5/gn :tinu( skcihc nrohgeL etihW dna nrohgeL etihW dezimatnaB ,matnaB fo spuorg lortnoc dna detaert latibrabonehp ni eneg nitca-β...

Ngày tải lên: 07/08/2014, 18:21

... three proteins; dots indicate amino acids identical between two proteins The small box indicates the position of substituted amino acids within the mEH catalytic triad (Glu in DmEH; Asp in Jheh1 ... precisely determined in this study Similar rapid and transient activations of self-defence protein genes in insects occur when insects are infected with microorganisms [35], in which case antimicrobial ... in wildtype and apterous mutant Drosophila Mol Cell Endocrinol 81, 205–216 23 Gilbert, L.I., Rybczynski, R & Tobe, S.S (1996) Endcrine cascade in insect metamorphosis In Metamorphosis (Girbert,...

Ngày tải lên: 07/03/2014, 21:20

... This finding supported our clinical testing regimen; all longitudinal frozen PBMC samples from each patient were tested in a single assay by a single operator, allowing assessment of vaccine potency ... defined as TIL cells which lack binding to negative tetramer and irrelevant tetramer and show specific binding to the relevant tetramer (TIL1520 binding to gp100 and TIL1143 binding to MART-1) ... (MART-1 control) working cell banks stored frozen in LN2 Freshly thawed longitudinal PBMC samples from each patient were analyzed for gp100 and MART-1 tetramer binding in a single assay using...

Ngày tải lên: 18/06/2014, 15:20

... combination of both methods greatly speeds up ascertaining when a NoV outbreak is occurring and which strain is responsible A SYBR green mastermix containing a proof reading enzyme would again ... panel are shown on the left binding to there respective probes Clinical samples binding to probe GII/4 can be seen on the right of both A and B Biotinylated RT- PCR Biotinylated reverse primers G1NVR ... Biotech (Ebersberg, Germany) were used in the following RT- PCR assay at a final volume of 50 µl The reaction contained µl of cDNA from the RT reaction, µl of 10× PCR Buffer, 1.5 mM MgCl2, µl of 10...

Ngày tải lên: 18/06/2014, 18:20

... combination of both methods greatly speeds up ascertaining when a NoV outbreak is occurring and which strain is responsible A SYBR green mastermix containing a proof reading enzyme would again ... panel are shown on the left binding to there respective probes Clinical samples binding to probe GII/4 can be seen on the right of both A and B Biotinylated RT- PCR Biotinylated reverse primers G1NVR ... Biotech (Ebersberg, Germany) were used in the following RT- PCR assay at a final volume of 50 µl The reaction contained µl of cDNA from the RT reaction, µl of 10× PCR Buffer, 1.5 mM MgCl2, µl of 10...

Ngày tải lên: 20/06/2014, 01:20

... of RT- PCR markers by lymph node status Lymph node involvement for all patients in the study had been assessed using haemotoxylin and eosin staining (Table 2) Certain patients had their sentinel ... purposes) ELF3 PCR - ve PCR - ve PCR - ve RT - ve Gen DNA cDNA RT +C P5 P 45 P 56 P 19 P 52 P 25 P 54 P 10 P 34 P8 Expression of RT- PCR markers by lymphovascular invasion (LVI) Fourteen patients ... (Figure 1) In samples containing cell per ml of blood (10 cells total), marker expression was detected in 2/3 samples indicating some loss of cells during the immunobead isolation In the main part of...

Ngày tải lên: 10/08/2014, 22:20

... 30 minutes at 37°C Direct in situ RT- PCR was carried out in a GeneAmp In Situ PCR System 1000 (Perkin Elmer Biosystems, Foster City, CA) using a GeneAmp Thermostable rTth RT- PCR kit (Perkin Elmer ... genome in formalin-fixed paraffin-embedded liver tissue by in situ reverse transcription polymerase chain reaction J Med Virol 1994, 44:406-409 Martinez A, Miller MJ, Quinn K, Unsworth EJ, Ebina ... limited proteinase K digestion; Step 2: amplification of reverse-transcribed cDNA using intron-spanning PCR primers (horizontal arrows) added in a single mix of reverse transcriptase, rTth polymerase...

Ngày tải lên: 11/08/2014, 00:21

... assay was initially highly successful in detecting H5N1 infections in Egypt [5,7] Since 2007, however, an increasing number of strains in Egypt escaping detection by this assay was reported [24] ... RT- qPCRs targeting the M gene of AIV and superior to a generic H5 RT- qPCR [23] when examining swabs which originated from experimental infections or from field samples of poultry holdings in Egypt ... testing, culling and vaccination of poultry is the key to mitigate the risk of human infections and fatalities in Egypt Controlling the endemic HPAIV H5N1 situation in Egypt is particularly painstaking...

Ngày tải lên: 12/08/2014, 01:22

... file 2) Miner software pinpoints the starting and ending points of PCR exponential phase from raw fluorescence data, and estimates primer set amplification efficiencies through a nonlinear regression ... staining The presence of contaminant DNA in the RNA samples was verified by PCR using primers spanning two exon and gel electrophoresis analysis No fragments of genomic DNA were identified in ... volume (20 μL) was incubated at 50°C for h following manufacturer’s instructions Inactivation of the reverse transcriptase was done by incubating the mixture at 70°C for 15 and the cDNA solution was...

Ngày tải lên: 12/08/2014, 03:21

... Discussion In this study, PVM was detected in several potato samples from an experimental farm in the province of Page of Figure Sensitivity of RT- PCR using primer set PVM3/PVM4 for detecting PVM in ... also tested in RT- PCR using this primer set to determine specificity None of them yielded any amplification products To evaluate the sensitivity of the RT- PCR for the detection of PVM in dormant ... designed and RT- PCR procedures were developed for the specific detection of PVM in various potato samples and for the confirmation of PCR amplicons The efficacy of RT- PCR for indexing seed potato...

Ngày tải lên: 12/08/2014, 04:21

... factors, subclinical infections in particular, might influence the outcome of vaccinations Eimeria species are regarded as ubiquitous parasites in most poultry environments, colonizing chicken ... decreases proportionally, increasing the risk of subclinical coccidiosis E tenella mainly replicates in the epithelium of the cecae, but developing stages of E tenella have been found in the bursa ... and during post mortem examination, eluted in sterile saline for one hour and kept frozen at -20°C until analysed All tissue samples and swab samples were initially analysed by Qiagen RT- PCR without...

Ngày tải lên: 12/08/2014, 18:21

... One-step RT- PCR for identification of contaminants in Kit I and Platinum Taq (A-C) One-step RT- PCR for identification of a contaminated component in Kit I The experiments were conducted in two independent ... They also used Invitrogen’s Superscript II RT and Platinum Taq for RTPCR, albeit in a two-step cDNA synthesis and PCR amplification procedure [13] As pointed out by Erlwein et al in the comments ... one-step RT- PCR reactions as indicated in Figure To examine whether the contaminant was RNA, μl of 10 μg/ml RNaseA (Cat no 19101) (QIAGEN) were added in the one-step RT- PCR reaction mixture as indicated...

Ngày tải lên: 13/08/2014, 01:20

... ca cỏc enzyme auxin-oxidase, iu ny cho thy nng auxin luụn cao hn nhng vựng tng hp chỳng ỉ Cytokinin Cytokinin (gm Kinetin, IBA, Zeatin v 2-iP) c phỏt hin sau auxin v gibberellin Ngi ta bit rng ... sung cytokinin s to iu kin thun li cho s phõn chia t bo v hỡnh thnh chi Cytokinin l cỏc hp cht adenin c thay th, cú nhúm cytokinin ni sinh c bit n l Zeatin v IBA, ngoi cũn cú nhúm cytokinin tng ... tớnh vi PCR Phỏt hin cỏc khim khuyt gene nh type cỏc mụ Phỏt hin cỏc vi sinh vt gõy bnh Recombinant PCR K thut footprinting DnaseI HLA ADN (cỏc khỏng nguyờn bch cu ngi) 2.2 Phng phỏp RT- PCR 22...

Ngày tải lên: 07/03/2016, 10:47

... bt nh Auxin cng c ch s phỏt trin ca chi nỏch v s hỡnh thnh phụi sinh dng mụi trng nuụi cy mụ so ỉ Cytokinin Cytokinin (gm Kinetin, IBA, Zeatin v 2-iP) c phỏt hin sau auxin v gibberellin Ngi ta ... sung cytokinin s to iu kin thun li cho s phõn chia t bo v hỡnh thnh chi Cytokinin l cỏc hp cht adenin c thay th, cú nhúm cytokinin ni sinh c bit n l Zeatin v IBA, ngoi cũn cú nhúm cytokinin tng ... vi PCR - Phỏt hin cỏc khim khuyt gene - nh type cỏc mụ - Phỏt hin cỏc vi sinh vt gõy bnh - Recombinant PCR - K thut footprinting DnaseI HLA ADN (cỏc khỏng nguyờn bch cu ngi) 2.2 Phng phỏp RT- PCR...

Ngày tải lên: 02/08/2016, 13:37

... 4.1.1 RT- PCR vi Taq beat hot start (Promega) 28 4.1.2 RT- PCR vi Taq polymerase ca Biorad v ABgene 29 4.2 Kt qu RT- PCR trờn mu RNA ly trớch 30 4.2.1 RT- PCR vi Taq beat hot start trờn ... cú th phỏt hin c bng RT- PCR ễng kt lun kt qu phn ng RT- PCR ph thuc rt nhiu vo mu phõn tớch (7).Wagstrom v cng s nghiờn cu ca mỡnh ó chng minh k thut RT- PCR trc tip cú kh nng phỏt hin RNA virus ... 3.2 Thnh phn phn ng RT- PCR 25 Bng 3.3 Chu k nhit cho phn ng RT- PCR 26 Hỡnh 4.1 Sn phm RT- PCR trờn mu RNA chun dựng vi Taq Beat Hot Start 28 Hỡnh 4.2 Sn phm RT- PCR trờn mu RNA chun...

Ngày tải lên: 29/10/2012, 14:23

... đến 18 acid amine gồm: glutamic acid, arginine, proline, aspatic acid, proline, tyrosine, lysine, aspatic acid, glutamic acid Ngoài ly trích đƣợc tinh dầu có mùi hƣơng nhẹ từ hạt tinh dầu đƣợc ... μl High wash stringency Thu RNA binding column Ly tâm/ 30 s 700 μl Low wash stringency Thu RNA binding column - Ly tâm/ 30 s - Ly tâm/ phút 80 μl Elution solution Thu RNA binding column - Ủ/ ... RNA binding column Ly tâm/ phút Thực tƣơng tự hết dung dịch tube hòa tan với EtOH 70 % 700 μl Low wash stringency Thu RNA binding column Ly tâm/ 30 s 80 μl dung dịch Dnase I Thu RNA binding column...

Ngày tải lên: 01/11/2012, 10:59

... Prawn which losses aquaculture industry in India The disease was reported in Guadeloupe and Martinique (India) subsequently in the five provinces of China The clinical sign of disease is whitish ... positive control from India was used We experimented successful process of Single-Step RT- PCR of Widada and Sahul Hameed The suitable condition for a Single-Step RT- PCR RT- PCR reaction is 1,5 mM ... thuật RT- PCR Nội dung nghiên cứu  Thử nghiệm qui trình Single - Step RT - PCR phát MrNV, XSV từ mẫu chứng dƣơng đƣợc cung cấp từ Ấn Độ  Khảo sát số điều kiện qui trình Single - Step RT - PCR...

Ngày tải lên: 05/11/2012, 14:01

... 4.1.1 RT- PCR vi Taq beat hot start (Promega) 28 4.1.2 RT- PCR vi Taq polymerase ca Biorad v ABgene 29 4.2 Kt qu RT- PCR trờn mu RNA ly trớch 30 4.2.1 RT- PCR vi Taq beat hot start trờn ... cú th phỏt hin c bng RT- PCR ễng kt lun kt qu phn ng RT- PCR ph thuc rt nhiu vo mu phõn tớch (7).Wagstrom v cng s nghiờn cu ca mỡnh ó chng minh k thut RT- PCR trc tip cú kh nng phỏt hin RNA virus ... 3.2 Thnh phn phn ng RT- PCR 25 Bng 3.3 Chu k nhit cho phn ng RT- PCR 26 Hỡnh 4.1 Sn phm RT- PCR trờn mu RNA chun dựng vi Taq Beat Hot Start 28 Hỡnh 4.2 Sn phm RT- PCR trờn mu RNA chun...

Ngày tải lên: 15/11/2012, 09:07

... town, Binh Duong province - In plants expressing disease symptom, the vascular bundles presented fluorescence in the phloem while symtomless plants not have fluorescence - Finding out the RT- PCR ... 27 2.4 Kỹ thuật PCR RT- PCR 2.4.1 PCR Kĩ thuật PCR PCR (Polymerase Chain Reaction- phản ứng tổng hợp dây chuyền nhờ polymerase) Kary Mullis cộng phát minh vào 1985 Đây cơng cụ sinh học phân tử, ... CƠNG NGHỆ SINH HỌC Giáo viên hƣớng dẫn: PGS TS BÙI CÁCH TUYẾN Sinh viên thực NGUYỄN MINH NAM Niên khóa: 2002 – 2006 Thành phố Hồ Chí Minh Tháng 09/2006 MINISTRY OF EDUCATION AND TRAINING NONG LAM...

Ngày tải lên: 17/11/2012, 09:44