amplification cloning and sequencing
Báo cáo y học: "Analysis of bacterial DNA in synovial tissue of Tunisian patients with reactive and undifferentiated arthritis by broad-range PCR, cloning and sequencing" pps
... procedures, and performed clinical and rheumatological data analyses AZ, CB and EC conducted assessment of Chlamydia trachomatis serology and DNA extraction BJ and JS participated in the design and coordination ... design and coordination of the study, and drafted the manuscript AH and AS analyzed microbiological and sequencing data, and revised the manuscript All authors read and approved the final manuscript ... PCR and/ or reverse transcription PCR systems to search for bacterial DNA and RNA in synovial samples from patients with various forms of arthritis, including ReA [12,14,17] By cloning and sequencing...
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Báo cáo y học: "Broad-range PCR, cloning and sequencing of the full 16S rRNA gene for detection of bacterial DNA in synovial fluid samples of Tunisian" pps
... AH and AS analyzed microbiological and sequencing data, and revised the manuscript All authors read and approved the final manuscript 15 16 17 Acknowledgements We thank Sebastien Chaussonneri and ... -80°C until analysis Automated DNA extraction from synovial fluid and broad-range PCR amplification of 16S rRNA genes, cloning and sequencing of bacterial DNA For 10 minutes, 500 μl of SF were centrifuged ... were found by sequencing short DNA fragments [12-14,16] Thus, the identification of such bacterial species after cloning and near-full length 16S DNA sequencing might be of interest and should be...
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Báo cáo khoa học: Purification, characterization, cDNA cloning and nucleotide sequencing of a cellulase from the yellow-spotted longicorn beetle, Psacothea hilaris ppt
... Proteins were visualized with CBB, cut out, and subjected to gas phase protein sequencing (model LF-3400 DT; Beckman) cDNA cloning, genomic PCR and sequencing A QuickPrepÒ Micro mRNA Purification ... without reducing agent and incubated at 33 °C for 30 to load an SDS/polyacrylamide gel The SDS/ PAGE analysis detected three proteins bands by CBB staining and one activity band by activity staining ... 1, standard sugars: G1, G2, G3, G4, cellopentaose (G5) and cellohexaose (G6) Lanes 2–5, G2–G5 treated with P hilaris cellulase at 37 °C for h and cellopentaose were degraded to cellobiose and...
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Báo cáo sinh học: " A general method for nested RT-PCR amplification and sequencing the complete HCV genotype 1 open reading frame" potx
... lanes and versus and 4) Random hexamers (Rndm) were more efficient than B4R1, a primer specific to the 3'-UTR (lanes and versus and 6, or lanes and versus and 10) For amplicon 2, AMVRT and M-MLV ... (lanes and versus and 8, or lanes and versus and 10) Lanes 11 and 12 are negative controls in which template RNA was omitted Reject primers Keep primers Figure Amplification optimization process Amplification ... degree of genomic heterogeneity and relatively low viral titres, efficient amplification and sequencing of the HCV ORF is possible We report optimized amplification and sequencing conditions for the...
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báo cáo hóa học:" A general method for nested RT-PCR amplification and sequencing the complete HCV genotype 1 open reading frame" pdf
... lanes and versus and 4) Random hexamers (Rndm) were more efficient than B4R1, a primer specific to the 3'-UTR (lanes and versus and 6, or lanes and versus and 10) For amplicon 2, AMVRT and M-MLV ... (lanes and versus and 8, or lanes and versus and 10) Lanes 11 and 12 are negative controls in which template RNA was omitted Reject primers Keep primers Figure Amplification optimization process Amplification ... degree of genomic heterogeneity and relatively low viral titres, efficient amplification and sequencing of the HCV ORF is possible We report optimized amplification and sequencing conditions for the...
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Tài liệu Soft Computing for Optimal Planning and Sequencing of Parallel Machining Operations docx
... study of the optimal planning and sequencing for parallel machining operations The combinatorial nature of sequencing and the complication of having precedence and mode constraints make the problem ... Nee and A.N Poo, Integrated application of expert systems and neural networks for machining operation sequencing, in Neural Networks in Manufacturing and Robotics, Y.C Shin, A.H Abodelmonem and ... industrial manufacturing, only a handful of papers are found on sequencing machining operations for parallel machines [3, 22] The combinatorial nature of sequencing and the complication of having...
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Tài liệu Báo cáo " synthesis, cloning and expression in escherichia coli of a gene coding for Mcoti-ii " ppt
... the 58 kD protein band was found in induced sample as the major band, acounted for 80% of total protein (lane 1,3,5 and 6) 58 kD Fig 10 SDS PAGE pattern of PI-17 proteins Mc Standard molecular ... the Future, Michal Lebl and Richard A Houghten (Editors), American Peptide Society, 2001, pp 387-388 Bozenna Rempola, T-Wilusz, W Markiewicz and M.Fkus, Synthesis, cloning and expression in E.coli ... Manual, University of Helsinki, Finland, (1998), pp 4-7 Hernandez, J.F., Gagnon, J., Chiche, L., Nguyen, T.M, Andrieu, J.P., Heitz, A., Trinh H.T., Pham T.T Chau., and Le Nguyen D., V Squash trypsin...
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Tài liệu Báo cáo khoa học: Molecular cloning and characterization of soybean protein disulfide isomerase family proteins with nonclassic active center motifs pdf
... (Fig 1D) GmPDIL-3a and GmPDIL-3b have two predicted thioredoxin domains between amino acids 65–164 and 403–481, and 68–167 and 406–508, respectively, corresponding to the a-domain and a¢-domain of ... (Fig 3A, lanes and 5) Anti-GmPDIL-3a serum reacted with both a 55 kDa and a 59 kDa band in western blot analysis of cotyledon proteins (Fig 3A, lane 3) Both the 55 and 59 kDa bands were N-glycosylated, ... left, and density (gÆmL)1) is indicated on the top (B) Microsomes were treated without (lanes and 2) or with (lanes and 4) proteinase K, in the absence (lanes and 3) or presence (lanes and 4)...
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Tài liệu Báo cáo khoa học: Cloning and characterization of CBL-CIPK signalling components from a legume (Pisum sativum) ppt
... treatment of pea seedlings were cold (A and B), salinity (C and D), wounding (E and F), drought (G and H), SA (I and J), ABA (K and L) and calcium (N and O) Panel M is the control for ABA responsive ... Figure 7A, D, I and L are diamidino-2phenylindole hydrochloride (DAPI) stained cells showing the nucleus, and Fig 7C, F, K and N are the merged images of A and B, D and E, I and J, and L and M, respectively ... 3) and a 26-kDa band (Fig 4G, lane 3), respectively In western blotting, the anti-PsCIPK and PsCBL antibodies detected PsCIPK as a single band of 58 kDa (Fig 4B, lane and 3, respectively) and...
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Tài liệu Báo cáo khóa học: Cloning and characterization of two distinct isoforms of rainbow trout heat shock factor 1 ppt
... amplification cycles of 0.5 at 94 °C, 0.5 at 50 °C and 0.5 at 72 °C, and one termination cycle of at 72 °C, with 35 cycles in total for HSF1a and HSF1b and 30 for heat shock cognate 70 (HSC70) Rainbow ... acids Ó FEBS 2004 By using 5¢- and 3¢-RACE, the full-length cDNAs of clones C1 and C2 without introns were determined to be 2083 bp and 2142 bp, respectively Clones C1 and C2 were Fig Comparison ... (rt) HSF1a and HSF1b with the sequences of zebrafish (z), chicken (c), mouse (m) and human (h) HSF1 The three domain structures, the DBD and the hydrophobic heptad repeats (HR-A/B and HR-C), are...
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Tài liệu Báo cáo khoa học: Tachykinin-related peptide precursors in two cockroach species Molecular cloning and peptide expression in brain neurons and intestine docx
... maderae and 15 in P americana Mass spectrometric analysis demonstrated the occurrence of the different TKRPs in glomeruli of the tritocerebrum and in cells of the optical lobe Results Cloning ... degenerated PCRs and RACE, we obtained full length preprotachykinin cDNA sequences for L maderae and P americana The cDNAs of 1512 bp 3366 R Predel et al (lem, GenBank accession number AY766011) and 1200 ... TKRPs are identical in L maderae and P americana, only six amino acid substitutions and one deletion were found (Fig 1) The signal peptide sequence (69.2% identity) and the spacer sequences between...
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Tài liệu Báo cáo Y học: Purification, characterization, cloning, and expression of the chicken liver ecto-ATP-diphosphohydrolase pot
... Ecto-Atpases and Related Ectonucleotidases (Vanduffel, L & Lemmens, R., eds), pp 1–8 Shaker Publishing B.V., Maastricht, the Netherlands Handa, M & Guidotti, G (1996) Purification and cloning of ... preparation of DNA for sequencing and transfection DNA sequencing was provided by the San Diego State University Microchemical Core Facility Transient and stable transfection COS-7 and HeLa cells were ... primers and thermal cycling conditions but with Advantage Taq DNA polymerase (Clontech) for subsequent TA cloning The 1.5-kb PCR product was gel purified and ligated to pCR2.1 (Invitrogen) and an...
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..GENE CLONING AND DNA ANALYSIS..GENE CLONING AND DNA ANALYSISAn IntroductionT.A. BROWNFaculty of Life Sciences University of Manchester ManchesterSixth EditionA John Wiley & Sons, Ltd., Publication. pdf
... from Cloned Genes 225 Gene Cloning and DNA Analysis in Medicine 245 Gene Cloning and DNA Analysis in Agriculture 264 Gene Cloning and DNA Analysis in Forensic Science and Archaeology 282 Glossary ... Gene Cloning and DNA Analysis in Research 163 10 Sequencing Genes and Genomes 165 10.1 The methodology for DNA sequencing 165 10.1.1 Chain termination DNA sequencing 166 Chain termination sequencing ... advent of gene cloning and the polymerase chain reaction What is gene cloning? What is PCR? Why gene cloning and PCR are so important 1.5.1 Obtaining a pure sample of a gene by cloning 1.5.2 PCR...
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Báo cáo khoa học: Molecular cloning and characterization of two soybean protein disulfide isomerases as molecular chaperones for seed storage proteins doc
... between a and b and b¢ and a¢ (Fig 1C) In the case of GmPDIL-2, Lys175, Glu43, Glu62, Glu68 and Glu135 were identified by N-terminal sequencing of the enzymatic digest fragments, respectively, and Glu541 ... and nitrogen for germination and early growth [18] Primary soybean seed storage proteins are globulins called glycinin and b-conglycinin They are folded and assemble into trimers in the ER, and ... b and b¢ and b¢ and a¢ in GmPDIL-2, the structure of these regions may be protease resistant The activity of recombinant GmPDIL-1 and GmPDIL-2, which catalyze oxidative refolding of reduced and...
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Báo cáo khoa học: Molecular cloning and characterization of methylenedioxy bridge-forming enzymes involved in stylopine biosynthesis in Eschscholzia californica doc
... japonica CYP719 was renamed CYP719A1, and EcCYPA and EcCYPB were designated CYP719A3 and CYP719A2, respectively Sequence analysis also showed that CYP719A2 and CYP719A3 had conserved eukaryotic ... examined whether CYP719A2 and CYP719A3 could function as CHS or STS Heterologous expression of CYP719A2 and CYP719A3 in S cerevisiae and their enzymological activities CYP719A2 and CYP719A3 were heterologously ... CYP719A3 converted (R,S)-cheilanthifoline and (S)-scoulerine to stylopine and nandinine at a comparable level (Fig 6) Furthermore, when (R,S)-cheilanthifoline and (S)-THC were used as mixed substrates...
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Báo cáo khoa học: Cloning and characterization of the genes encoding toxic lectins in mistletoe (Viscum album L) pot
... variants giving 169 and 246 bp fragments SalI cuts the amplification product of ml2p, giving 66, 121 and 243 bp fragments, and the amlpification product of ml3p and ml3.1p, giving 50 and 365 bp fragments ... were purified from agarose gel (GFX PCR DNA and Gel Band Purification Kit, USB) and cloned into pGEM7zf(+) or pGEM3zf(+) vector (Promega) DNA isolation and Southern blot analysis Whole genomic DNA ... separated on 2% agarose gel and the bands corresponding to genomic fragment (length of 415/430 bp) and competitor fragment (length of 388 bp) were excised Radioactivity of the bands was then measured...
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Báo cáo khoa học: Cloning and expression of the first nonmammalian interleukin-11 gene in rainbow trout Oncorhynchus mykiss pdf
... clone as well as a genomic DNA clone were isolated and sequenced, and the expression and modulation of this molecule was studied Results Cloning and characterization of rainbow trout IL-11 A cDNA ... bold and italics The ATTTA motifs in the 5¢- and 3¢-UTRs, the start and stop codons, the signal peptide predicted by SIGNALP 3.0 [23] and the potential N-glycosylation site (NQT) are in bold and ... healthy rainbow trout and used for amplification of IL-11 (Primer EF and ER, expected size: 271 bp) and b-actin (expected size: 276 bp) The cycle numbers were 21 and 32 for b-actin and IL-11, respectively...
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Báo cáo Y học: Cloning and expression of sterol D14-reductase from bovine liver potx
... (DMEM), and foetal bovine serum were purchased from Gibco-BRL (Milan, Italy) TOPO -cloning kit was from Invitrogen (Leek, the Netherlands) RNAse inhibitor was from Ambion (Austin, TX, USA) The Expand ... First-strand synthesis was performed at 42 °C for 45 and then the enzyme was inactivated at 90 °C for Following ®rststrand synthesis, PCR of D14-SR cDNA was carried out using appropriate primers and ... Saccharomyces cerevisiae D14-SR (55%) (Fig 1), and other sterol reductases [50% and 49% similarity to human and A thaliana sterol D7-reductases, respectively, and 44% to S cerevisiae sterol D24(28)-reductase]...
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Báo cáo Y học: Molecular cloning and characterization of isomultiflorenol synthase, a new triterpene synthase from Luffa cylindrica, involved in biosynthesis of bryonolic acid ppt
... at days 16 and 20 and low at days and The induction of this mRNA preceded the accumulation of bryonolic acid In contrast, the level of cycloartenol synthase mRNA was high at days and and decreased ... plasmid for yeast, Kpn I and Xba I sites were introduced into the 50 and 30 -termini of the deduced ORF of LcIMS1 cDNA by PCR (25 cycles with 40 s at 94 8C, 40 s at 50 8C and at 72 8C) with Pfu ... ATTG-30 (Kpn I site underlined) and 50 -CCTCTAGAGTA AAAGTCTCCAATC-30 (Xba I site underlined) were used to modify the cDNA The PCR product was digested with Kpn I and Xba I, and ligated to the sites...
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Báo cáo khoa học: ˚ cDNA cloning and 1.75 A crystal structure determination of PPL2, an endochitinase and N-acetylglucosaminebinding hemagglutinin from Parkia platycephala seeds potx
... conserved strands b3 and b4 (Fig 4A, boxed), and the hydrogen bond network between residues Asp120 and Gly121 and Val74 (Fig 4A,B) [33], are also conserved in PPL2 On the other hand, the largest ... bond, and Asp125 and Tyr183 would contribute to the stabilization of the oxazolinium intermediate [45] In PPL2, these residues correspond to Asp125, Glu127 and Tyr182 (Figs and 4A) Asp125 and Glu127 ... the combination of N-terminal sequencing and cDNA cloning contains 271 amino acid residues, including the six conserved cysteine residues of class III chitinases, and the putative catalytic residues...
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