extraction reverse transcriptase pcr and cycle sequencing
Báo cáo sinh học: " Genetic characterisation of the recent foot-and-mouth disease virus subtype A/IRN/2005" pptx
... phylogenies of 2B and 3AB and in addition with the Panasia lineage of type O, an Asia1 and O5 isolate from India and the A Iran105 isolate from Iran A relationship between Asia1 and the Panasia ... reaction as follows: 95°C for 15 min, and five cycles of 94°C for 30 sec, 59°C for and 72°C for 30 sec, and then 35 cycles of 94°C for 30 sec, 61°C for 30 sec and 72°C for 30 sec, followed by a final ... information PN helped in finding primers and optimizing the PCR SA is project coordinator and conceived the study and helped to draft the manuscript All authors read and approved the final manuscript...
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Báo cáo hóa học: " Genetic characterisation of the recent foot-and-mouth disease virus subtype A/IRN/2005" ppt
... phylogenies of 2B and 3AB and in addition with the Panasia lineage of type O, an Asia1 and O5 isolate from India and the A Iran105 isolate from Iran A relationship between Asia1 and the Panasia ... reaction as follows: 95°C for 15 min, and five cycles of 94°C for 30 sec, 59°C for and 72°C for 30 sec, and then 35 cycles of 94°C for 30 sec, 61°C for 30 sec and 72°C for 30 sec, followed by a final ... information PN helped in finding primers and optimizing the PCR SA is project coordinator and conceived the study and helped to draft the manuscript All authors read and approved the final manuscript...
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Báo cáo y học: " Dideoxynucleoside HIV reverse transcriptase inhibitors and drug-related hepatotoxicity: a case report" pot
... data and revised the manuscript LB and MG analyzed the liver biopsy and revised the manuscript CT conceived the study and participated in the drafting of the manuscript All authors read and approved ... contributions GL conceived the study and participated in drafting the manuscript II participated in the study design and in drafting the manuscript SCL, GC and SC participated in conceiving the ... determinations in June and October 2003 Moreover, HCV-RNA was repeated at the time of HT and still resulted negative Further, hepatitis A virus immunoglobulin M (HAV-IgM) resulted negative and the patient...
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Báo cáo khoa học: Alizarine derivatives as new dual inhibitors of the HIV-1 reverse transcriptase-associated DNA polymerase and RNase H activities effective also on the RNase H activity of non-nucleoside resistant reverse transcriptases pot
... both RNA and DNA dependent; and (b) RNase H activity that selectively hydrolyzes the RNA strand of the RNA:DNA hybrid formed during synthesis of the minus () strand DNA that uses (+)-strand RNA ... HIV-1 reverse transcriptase induced by nonnucleoside reverse transcriptase inhibitor binding Curr HIV Res 2, 323332 30 Pata JD, Stirtan WG, Goldstein SW & Steitz TA (2004) Structure of HIV-1 reverse ... without RT; lane 2, plus RT, lanes 38, plus RT and K-49 (100, 33, 11, 3.3, 1.1 and 0.33 lM); lanes 914, plus RT and RDS 1634 (100, 33, 11, 3.3, 1.1 and 0.33 lM) (C) LineweaverBurk plot of the inhibition...
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Báo cáo Y học: The processivity and fidelity of DNA synthesis exhibited by the reverse transcriptase of bovine leukemia virus pot
... Comparison of HIV-1 and avian myeloblastosis virus reverse transcriptase fidelity on RNA and DNA templates J Biol Chem 264, 10888–10896 31 Richetti, M & Buc, H (1990) Reverse transcriptases and genomic ... (1992) Retroviral reverse transcription and integration: progress and problems Annu Rev Cell Biol 8, 275–306 29 Bakhanashvili, M & Hizi, A (1992) The fidelity of the reverse transcriptase of human ... analysed and described in Materials and methods The apparent Km and Vmax values were determined from at least two independent experiments performed as described in Materials and methods and in...
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Báo cáo sinh học: " Development of a real-time RT-PCR and Reverse Line probe Hybridisation assay for the routine detection and genotyping of Noroviruses in Ireland" pdf
... utilised methods are Electron Microscopy (E.M) and Reverse Transcription Polymerase Chain Reaction (RT -PCR) RT -PCR is the preferred method as it is rapid and very sensitive; however, it relies heavily ... containing GI/2 and GII/4 PCR products corresponding to Southampton virus and Oxford B2S16 Detection levels of these plasmid molecules were 107 to 101 for GI and × 107 to × 101 for GII (Fig 1A and Fig ... Journal 2007, 4:86 http://www.virologyj.com/content/4/1/86 Table 1: Primers and probes used for Lightcycler RT -PCR and Reverse Line Blot hybridization assay Primers Primer Sequence Reference COG1F...
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Báo cáo sinh học: " Quantitative assessment of the effect of uracil-DNA glycosylase on amplicon DNA degradation and RNA amplification in reverse transcription-PCR" pdf
... generated by standard PCR and RT -PCR assays In addition to rapid quantification of template RNA, real-time PCR and RTPCR offers significant advantages over standard PCR and RT -PCR for detection ... samples containing UNG and incubated at 30°C for 30 prior to RT -PCR was 1.56 cycles, a value in agreement with results shown above Discussion The techniques of PCR and RT -PCR offer several advantages ... occurs The cycle at which a positive reaction is first detectible, termed the cycle threshold (CT) is proportionate to the concentration of template in a sample Real-time PCR and RT -PCR amplification...
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Báo cáo hóa học: " Development of a real-time RT-PCR and Reverse Line probe Hybridisation assay for the routine detection and genotyping of Noroviruses in Ireland" ppt
... utilised methods are Electron Microscopy (E.M) and Reverse Transcription Polymerase Chain Reaction (RT -PCR) RT -PCR is the preferred method as it is rapid and very sensitive; however, it relies heavily ... containing GI/2 and GII/4 PCR products corresponding to Southampton virus and Oxford B2S16 Detection levels of these plasmid molecules were 107 to 101 for GI and × 107 to × 101 for GII (Fig 1A and Fig ... Journal 2007, 4:86 http://www.virologyj.com/content/4/1/86 Table 1: Primers and probes used for Lightcycler RT -PCR and Reverse Line Blot hybridization assay Primers Primer Sequence Reference COG1F...
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Báo cáo hóa học: " Sargassum fusiforme fraction is a potent and specific inhibitor of HIV-1 fusion and reverse transcriptase" pot
... http://www.virologyj.com/content/5/1/8 Figure Inhibition of X4 and R5-tropic HIV-1 Inhibition of X4 and R5-tropic HIV-1 GHOST X4/R5 and GFP expressing cells were plate at × 105/well in 12-well plates and incubated at 37°C in CO2 ... demonstrating post entry inhibition of viral life cycle (Fig 5A) First step after HIV-1 entry is reverse transcription and cDNA formation by viral RT, and therefore we next investigated for possible ... with the viral CD4 receptor and direct inhibition of reverse transcriptase Conclusion Taken together, we have demonstrated an average of 53% inhibition of HIV-1 fusion, and approximately 47% of...
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báo cáo hóa học:" Quantitative assessment of the effect of uracil-DNA glycosylase on amplicon DNA degradation and RNA amplification in reverse transcription-PCR" ppt
... generated by standard PCR and RT -PCR assays In addition to rapid quantification of template RNA, real-time PCR and RTPCR offers significant advantages over standard PCR and RT -PCR for detection ... samples containing UNG and incubated at 30°C for 30 prior to RT -PCR was 1.56 cycles, a value in agreement with results shown above Discussion The techniques of PCR and RT -PCR offer several advantages ... occurs The cycle at which a positive reaction is first detectible, termed the cycle threshold (CT) is proportionate to the concentration of template in a sample Real-time PCR and RT -PCR amplification...
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báo cáo hóa học:" Substitutions in the Reverse Transcriptase and Protease Genes of HIV-1 Subtype B in Untreated Individuals and Patients Treated With Antiretroviral Drugs" potx
... copies/mL and in whom genotypic analysis was performed (n = 51) Plasma was obtained during 20002001 All subjects harbored subtype-B HIV-1 viruses and provided informed consent Sequencing of the RT and ... extracted using the QIAamp kit and RNA products were amplified by polymerase chain reaction (PCR) as described.[9] The sequencing of DNA products was carried out by standard methodology using kits ... human immunodeficiency virus type reverse transcriptase Nucleic Acids Res 1998, 26:3066-3072 Abstract Roberts J, Bebenek K, Kunkel T: The accuracy of reverse transcriptase from HIV-1 Science 1998,...
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Báo cáo y học: "Expression analysis of three isoforms of hyaluronan synthase and hyaluronidase in the synovium of knees in osteoarthritis and rheumatoid arthritis by quantitative real-time reverse transcriptase polymerase chain reaction" potx
... -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 ... 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...
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Báo cáo y học: ": Prevalence of reverse transcriptase and protease mutations associated with antiretroviral drug resistance among drug-naïve HIV-1 infected pregnant women in Kagera and Kilimanjaro regions, Tanzania" pptx
... Triomune, a combination of two nucleoside reverse transcriptase inhibitors (NRTI) Stavudine (d4T) and Lamivudine (3TC), and a non-nucleoside reverse transcriptase inhibitor (NNRTI) Nevirapine ... compliance with institutional policies and national guidelines Viral load, RNA extraction, RT PCR, secondary PCR and sequencing Viral load was determined in ml of plasma using COBAS AmpliPrep (ROCHE ... in Uganda J Acquir Immune Defic Syndr 2001, 26(5):495-500 Galluzzo CM: Antiretroviral Resistance Mutations in Untreated Pregnant Women with HIV Infection in Uganda and Rwanda AIDS Research and...
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Báo cáo y học: " Efficacy, safety and pharmacokinetic of once-daily boosted saquinavir (1500/100 mg) together with 2 nucleos(t)ide reverse transcriptase inhibitors in real life: a multicentre prospective study" ppt
... resistance (L90M) and were excluded from further analysis Among the remaining cases, 33 (27.5%) had wild-type isolates, and 71 (59.1%) had resistance mutations in the reverse transcriptase (TAMs ... with and without chronic hepatitis and/ or cirrhosis, only (0.6%) and 13 cases (5.4%), respectively, developed grade 3-4 transaminase increases Moreover, none of these cases was symptomatic, and ... safety and pharmacokinetic of once-daily boosted saquinavir (1500/100 mg) together with nucleos(t)ide reverse transcriptase inhibitors in real life: a multicentre prospective study AIDS Research and...
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Báo cáo y học: " A Leu to Ile but not Leu to Val change at HIV-1 reverse transcriptase codon 74 in the background of K65R mutation leads to an increased processivity of K65R+L74I enzyme and a replication competent virus" pptx
... antagonism between K65R and thymidine analog mutations in HIV-1 reverse transcriptase AIDS 2006, 21:1405-1414 Rhee SY, Liu TF, Holmes SP, Shafer RW: HIV-1 subtype B proteases and reverse transcriptases ... bottom six (1-6) bands between WT and K65R+L74V RTs (p = 0.38) and WT and K65R+L74I (p = 0.49) by paired student t-test analysis However, a significant increase in the densities of bands 25-30 was ... cDNA bands 16 (p = 0.384) and bands 7-12 (p = 0.237) However significant increase in the densities for larger bands (13-36) synthesized with K65R+L74I RT was obtained The p values were 0.016 (bands...
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Báo cáo y học: " A duplex real-time reverse transcriptase polymerase chain reaction assay for detecting western equine and eastern equine encephalitis viruses?" docx
... 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 ... Lambert et al 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 ... developed and validated a duplex realtime RT -PCR system for both WEEV and EEEV detection at the same time The primers and probes were selected from E3 gene for WEEV and NSP3 gene for EEEV, and the...
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Báo cáo y học: " The RNA binding protein HuR does not interact directly with HIV-1 reverse transcriptase and does not affect reverse transcription in vitro" doc
... Figure Purification and biophysical characterization of GSTand NusA-HuR (A) Analysis of the expression and purification of GSTHuR by SDS PAGE Lanes and illustrate total protein and total soluble ... immunodeficiency virus type reverse transcriptase Methods Enzymol 1995, 262:130-144 34 Le Grice SF, Gruninger-Leitch F: Rapid purification of homodimer and heterodimer HIV-1 reverse transcriptase by metal ... the atomic level and to elucidate the mechanism(s) by which HuR influences HIV-1 reverse transcription activity in vitro, we prepared and characterized four HuR protein constructs and investigated...
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Báo cáo y học: "Comparative biochemical analysis of recombinant reverse transcriptase enzymes of HIV-1 subtype B and subtype " ppt
... 3TC-TP, and TFV-DP, or NNRTIs NVP, EFV, and ETR The results of Table show that both the subtype B and the two subtype C RT enzymes from isolates BG05 and M01 (GenBank accession number AF492609 and ... Inc Efaverenz (EFV) and nevirapine (NVP) were obtained from BristolMyers Squibb Inc and Boehringer Ingelheim Inc, respectively Recombinant reverse transcriptase expression and purification The ... Sarafianos SG, Marchand B, Das K, Himmel DM, Parniak MA, Hughes SH, Arnold E: Structure and function of HIV-1 reverse transcriptase: molecular mechanisms of polymerization and inhibition J Mol...
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Báo cáo y học: "The evolution of HIV-1 reverse transcriptase in route to acquisition of Q151M multi-drug resistance is complex and involves mutations in multiple domains" ppt
... identified by agarose gel electrophoresis and purified using illustra GFX PCR DNA and Gel Band Purification Kit (GE Healthcare), and sequenced by the dideoxy ABI sequencing systems in both directions ... CHAP2 second-line NRTIs ddI and ABC showed a cumulative decrease in susceptibility in the order; 1. 2and 1.7-fold at months, 4- and 6-fold at 17 months, and finally 9.9- and 10.8-fold at 37 months, ... Switzerland for pCMVΔ8.91 and pMDG; and Thumbi Ndung’u, Boris Renjifo and Max Essex for p8MJ4 We also thank Soo-Yoon Rhee, Stanford University HIV database for help with database sequence analysis and...
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