samples cell lines and dna extraction
Báo cáo y học: " Revisiting HIV-1 uncoating" doc
Ngày tải lên :
13/08/2014, 01:20
... (IN), Vpr) and numerous cellular proteins, such as Cyclophilin A and APOBEC3G [9] Its main function is to organise and contain the viral genome for optimal delivery in target cells and efficient ... transcription involves firstly the formation of the minus strand strong-stop DNA, a strand transfer event, and the synthesis of the minus strand DNA with concomitant degradation of the RNA template In ... PPT (cPPT) and 3’ PPT, resist degradation by RNase H and serve as primers for synthesis of plus-strand DNA [49,53,54] Reverse transcription proceeds with synthesis of plus-strand DNA, involves...
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Nghiên cứu xây dựng quy trình phát hiện nhanh virus cúm gia cầm a h5n1 trong mẫu bệnh phẩm bằng kỹ thuật multiplex reverse transcription polymerase chain reaction
Ngày tải lên :
10/02/2014, 20:39
... (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, ... “Glycosylation of haemagglutinin and stalk-length of neuraminidase combine to regulate the growth of avian influenza viruses in tissue culture” Virus Res, 79(1-2): 177-185 Beare AS and Webster RG (1991),...
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Báo cáo khoa học: Ribonuclease H: properties, substrate specificity and roles in retroviral reverse transcription ppt
Ngày tải lên :
30/03/2014, 02:20
... which one strand is entirely DNA and the other strand is RNA at the 5¢-end followed by a stretch of DNA, the HIV-1 and M-MLV retroviral RNases H strongly prefer to cleave the RNA strand one nucleotide ... a recessed DNA 3¢-end DNA 3¢-end-directed cleavage A recessed DNA 3¢-end in a DNA ⁄ RNA hybrid is recognized by the polymerase activity of reverse transcriptase as a primer terminus and is utilized ... of the substrate Fig Three cleavage modes for retroviral RNases H DNA ⁄ RNA hybrids are drawn with RNA strands in red and DNA strands in black In the internal cleavage mode, the arrows mark the...
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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
Ngày tải lên :
19/06/2014, 08:20
... real-time PCR and RTPCR offers significant advantages over standard PCR and RT-PCR for detection of DNA or RNA in terms of reduced sample handling, the time required for analysis and analytical ... 17.09 21.48 DNA (1:107)b 24.89 DNA (1:108) 28.22 DNA (1:109) 31.75 DNA (1:1010) Mean CT increase for RNA Mean CT increase for DNA UNG concentration was 0.25 units per 25 µl reaction and incubation ... 14.47 21.46 DNA (1:107)b 24.73 DNA (1:108) 28.08 DNA (1:109) 31.14 DNA (1:1010) Mean CT increase for RNA Mean CT increase for DNA UNG concentration was 0.25 units per 25 µl reaction and incubation...
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Báo cáo hóa học: " A broad spectrum, one-step reverse-transcription PCR amplification of the neuraminidase gene from multiple subtypes of influenza A virus" pdf
Ngày tải lên :
20/06/2014, 01:20
... in the extraction of RNA and edited the manuscript along with EV, WC, and CB EV was also involved with virus culture, RNA extraction and RT-PCR validation WC and CB provided clinical samples, ... (description data not shown); M, 100 bp DNA Hyperladder II (Bioline); and negative control (- ctrl: water instead of template) Samples and RNA extraction Animal samples from allantoic fluids Virus ... freeze stored influenza A samples and these were mostly N2 subtype (see Table and Figure 4) From these influenza A clinical NPAs samples, two of them labeled as #17 and # 31 in Table were initially...
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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
Ngày tải lên :
20/06/2014, 04:20
... real-time PCR and RTPCR offers significant advantages over standard PCR and RT-PCR for detection of DNA or RNA in terms of reduced sample handling, the time required for analysis and analytical ... 17.09 21.48 DNA (1:107)b 24.89 DNA (1:108) 28.22 DNA (1:109) 31.75 DNA (1:1010) Mean CT increase for RNA Mean CT increase for DNA UNG concentration was 0.25 units per 25 µl reaction and incubation ... 14.47 21.46 DNA (1:107)b 24.73 DNA (1:108) 28.08 DNA (1:109) 31.14 DNA (1:1010) Mean CT increase for RNA Mean CT increase for DNA UNG concentration was 0.25 units per 25 µl reaction and incubation...
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Báo cáo hóa học: "Rapid detection of sacbrood virus (SBV) by one-step reverse transcription loop-mediated isothermal amplification assay" pptx
Ngày tải lên :
21/06/2014, 19:20
... contributions JY, YR and KS carried out most of the experiments and wrote the manuscript, and should be considered as first authors XP and ZL critically revised the manuscript and the experiment ... 63°C, 64°C and 65°C The DNA products from all reactions except that done at 59°C showed an obvious ladder-like pattern on the gel The intensities of the DNA products at 63°C, 64°C and 65°C were ... from Chongqing Academy of Animal Science All samples were stored at −20°C before RNA extraction Total RNA was extracted from SBV isolates using an RNA extraction kit (TaKaRa Biotechnology, Dalian,...
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Báo cáo khoa học: "TaqMan reverse transcription polymerase chain reaction for the detection of Japanese encephalitis virus" pot
Ngày tải lên :
07/08/2014, 18:20
... for detection of JEV by real-time RT-PCR The KV1899 and Anyang 300 strains were cultured in TF104 cells (a cell line cloned from MA104 cells), and the Nakayama strain was propagated from the infected ... in TF104 cell and Anyang 300 strain, which was an attenuated virus) From days post inoculation, blood samples from pigs inoculated with JEV were collected and plasmas were separated and stored ... primer (20 pM), µl of DEPC water and 15 µl of cDNA After incubation at 95oC, the cDNA was amplified by 45 threestep cycles: 10 s at 95oC, 20 s at 55oC and 20 s at 72oC and at 72oC for final extension...
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Báo cáo khoa học: "Development of a Lightcycler-based reverse transcription polymerase chain reaction for the detection of foot-and-mouth disease virus" pps
Ngày tải lên :
07/08/2014, 18:21
... )sgip dna swoc( slamina morf detcelloc erew IPD dna IPD neewteb detcelloc selpmas gnaborP ehT deraperp erew 2002 ni kaerbtuo DMF eht gnirud smraf gip 41 morf detcelloc seussit lailehtipe dna sdiulf ... noissucsiD IPD dna IPD neewteb RCP-TR lanoitnevnoc yb detceted eb dluoc ANR lariv dna suriv evil htob ,revewoH IPD.3 dna IPD neewteb ANR lariv rof evitisop erew sgip dna ,IPD dna IPD neewteb ... deniatbo erew VDMF rof sevruc dradnatS T T T T 05 05 stes eborp/remirp etauqeda fo noitceles dna seires noitulid dradnats eht fo RCP-TR T/R stluseR 05 05 05 ASILE-gA dna RCP-TR lanoitnevnoc fo esoht...
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Báo cáo y học: "Investigation of infectious agents associated with arthritis by reverse transcription PCR of bacterial rRNA" ppsx
Ngày tải lên :
09/08/2014, 01:21
... ensure that no contamination of samples occurred during protocol implementation Bacterial ribosomal DNA fragments were amplified from total cDNA and bacterial genomic DNA by PCR amplification using ... primers R1 and R2, as previously described in detail [14] All PCR negative samples were tested for potential PCR inhibitors by the addition of ng Escherichia coli genomic DNA, to their cDNA, and retested ... was extracted from the SF samples and approximately half was then treated with DNase I, and then aliquots of both were subjected to RT-PCR using universal primers R1 and R2 for detection of bacterial...
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Báo cáo khoa hoc:" Quantitative competitive reverse transcription polymerase chain reaction is not a useful method for quantification of CD4 and CD8 cell status during HIV infection" pptx
Ngày tải lên :
11/08/2014, 08:20
... 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 expressing ... CD4:CD8 cell ratios and are therefore likely to be HIV infected (the majority of the samples sent for flow cytometry are HIV-positive) The relationship between the mRNA and cell number for CD4 and ... between CD4 and CD8 mRNA levels and numbers of cells expressing these proteins on their surface The formation of CD4gp160 complexes in intracellular compartments may contribute to cell surface...
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Báo cáo y học: " Variability in a dominant block to SIV early reverse transcription in rhesus monkey cells predicts in vivo viral replication and time to death" pot
Ngày tải lên :
12/08/2014, 04:20
... 0.6209 17.5 % SIVsmE543-GFP+ B-LCL B Sensitive cell Resistant cell Sensitive cell + Resistant cell Sensitive/Sensitive cell Sensitive/Resistant cell 15.0 12.5 10.0 7.5 5.0 2.5 0.0 10 20 30 40 ... rhesus monkey PBMC, with susceptible cell lines supporting robust viral replication and resistant cells supporting little to no viral replication (Fig 1C and 1D) Rhesus monkey PBMC that exhibited ... cycle VSV-G pseudotyped SIVSsm543-GFP, and a significant correlation was observed for each cell population between % SIVsm543-GFP+ cells and the TCID50 of these cells for wild type SIVsmE660 (Figure...
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Báo cáo y học: " A multiplex reverse transcription-nested polymerase chain reaction for detection and differentiation of wild-type and vaccine strains of canine distemper virus" pptx
Ngày tải lên :
12/08/2014, 04:20
... 30 samples in Heilongjiang and Jilin provinces, and Fifty-one field samples from dogs, raccoons, foxes, and minks in Shandong provinces were assayed by RT-nPCR; the background of the 51 field samples ... cells infected with CDV vaccine strain, wild-type strain, mixed CDV vaccine and wild-type strains, field samples from Heilongjiang and Jilin province, CPV, CAV, CCV, RV, NDV, and uninfected cells ... improved[15] Complementary DNA and RNA probes have been used to detect RNA and mRNA of the CDV genome with improved specificity and sensitivity[6,7] Primers P1 and P4 (specific for CDV and conserved among...
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Retrovirology Review BioMed Central Open Access When is it time for reverse transcription to docx
Ngày tải lên :
12/08/2014, 23:20
... basic residues and zinc fingers), (iv) the primer tRNALys3 bound to RTp66-p51(in blue), (v) the viral RNA and RT, (vi) NC and RT and (vii) NC and NC molecules Viral proteins such as IN and Vif that ... RNA) (B: DNA) Nucleic acid copy numbers in virions (100ng p24) 108 wt 1ZF1 1ZF2 1ZF1ZF2 107 106 105 104 103 102 FL RNA MS RNAs ssDNA GagDNA FL DNA MS cDNAs Figure acids content of wild-type and NC ... minus-strand DNA transfer by nucleocapsid protein during reverse transcription of the retroviral genome EMBO J 1994, 13(4):973-81 Tsuchihashi Z, Brown PO: DNA strand exchange and selective DNA annealing...
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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
Ngày tải lên :
12/08/2014, 23:23
... 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 ... Conceived and designed the experiments: JA, IJLB, AMG and NSC Performed the experiments: JA, IJLB, SD, KH and JC Analyzed the data: JA, IJLB, AMG and NSC Wrote the paper: JA, IJLB, AMG and NSC ... unstable after purification and underwent substantial degradation at room temperature (Fig 2A) In contrast, the NusA-HuR fusion was stable and was used in the NMR and HIV-1 RT DNA synthesis reactions...
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Báo cáo y học: "Subtype-associated differences in HIV-1 reverse transcription affect the viral replication" ppt
Ngày tải lên :
13/08/2014, 01:20
... to decreased cDNA accumulation in reverse transcription complexes Accumulation of strong-stop (A and C) and positive-strand (B and D) viral DNA in RTCs isolated at and h p.i SupT1 cells were synchronously ... used for isolation of total cellular DNA Cell viability was measured using Vi -Cell cell viability analyzer (Beckman Coulter) Cellular DNA was purified with IsoQuick DNA Isolation kit for sequence ... quantitative analysis of cDNA and proviral DNA, total DNA was purified using the IsoQuick DNA Isolation kit and then analyzed by real-time quantitative PCR Fractionation of cells and isolation of the...
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Báo cáo y học: " Blocking premature reverse transcription fails to rescue the HIV-1 nucleocapsid-mutant replication defect" ppsx
Ngày tải lên :
13/08/2014, 01:20
... cells were infected with PEG-precipitated virus for h and cells were harvested at the indicated time points DNA from infected cells was isolated using a Qiagen DNA Blood Mini-Kit, and viral and ... cotransfected cells with different ratios of NC WT and NC mutant proviral plasmids and examined the virus for infectivity in TZM-bl cells and also measured quantities of intravirion DNA Figure 7A ... with DNase I and subtilisin to remove extra-virion contaminating plasmid DNA (Figure 1, right), and measured intravirion DNA by quantitative PCR (qPCR) to assess the levels of minus-strand strong-stop...
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Báo cáo y học: "Isolated HIV-1 core is active for reverse transcription" pptx
Ngày tải lên :
13/08/2014, 05:22
... first-strand transfer, and fulllength minus strand synthesis was also detectable above background (Fig 1B) However, the signal was not sufficient to determine whether products indicating secondstrand ... 1C–F), and fractions were then analyzed by western analysis using purified anti-HIV-1 IgG (NIH AIDS Research and Reference Reagent Program) Multiple protein bands in the peak virus fractions and ... making DNase I-resistant full length minus-strand DNA, albeit inefficiently, without requiring capsid release Our isolation of active cores without detergent treatment was fortuitous and reproducible;...
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Báo cáo y học: "Early and transient reverse transcription during primary deltaretroviral infection of sheep" ppt
Ngày tải lên :
13/08/2014, 06:20
... the provirus and the 3' cellular flanking sequence Lines and represent the first two mitoses of the infected cells harboring the integrated provirus For each cell, the RU5 sequence and the 3' flanking ... Deltaretroviruses include human T -cell leukemia viruses type -1 [7] and -2 (HTLV-1 and 2) [8], the recently discovered HTLV-3 [9] and -4 [10], simian T -cell leukemia viruses (STLV) [11], and the bovine leukemia ... that interfere with many host cell pathways including cell cycle, apoptosis and DNA repair [2,3] This results in the persistent clonal expansion of infected cells and generates a significant level...
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Báo cáo y học: " Contribution of the C-terminal tri-lysine regions of human immunodeficiency virus type 1 integrase for efficient reverse transcription and viral DNA nuclear import" pps
Ngày tải lên :
13/08/2014, 09:21
... 2; and 4; and 10) and 5D) For KK240,4AE mutant, approximately 51% of viral DNA was nucleusassociated (Fig 5C (upper panel, lane and 8) and 5D) Remarkably, in KK215,9AA infected sample, viral cDNA ... 106 cells were lysed and serial-diluted cell lysates were analyzed by two-step Alu-PCR and Southern blot for specific detection of integrated proviral DNA from infected cells (Upper panel) The DNA ... purposes) Retrovirology 2005, 2:62 Cell transfection and immunofluorescence assay DNA transfection in 293T and HeLa cells were performed with standard calcium phosphate DNA precipitation method For...
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