fi cation pcr2 and pcr3
Tài liệu Báo cáo khoa học: Identification, sequencing, and localization of a new carbonic anhydrase transcript from the hydrothermal vent tubeworm Riftia pachyptila docx
... n=4 RpCAbr amplification normalized with 18S amplification RpCAtr amplification normalized with 18S amplification Fig Normalized amplifications of RpCAbr and RpCAtr with 18S ampli cation The number ... transcriptase was added (1 lL) to each reaction mixture and all reactions were incubated at 42 °C for h and finally placed on ice Ampli cation Specific pairs of CA primers (Table 1) located in the 3¢ ... triplicates Ampli cation conditions were 40 cycles with the following profile: 95 °C for 30 s, 60 °C for 30 s, and 72 °C for For each kind of tissue, standard curves were generated for 18S and the CA...
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Báo cáo khoa học: Biochemical characterization of USP7 reveals post-translational modification sites and structural requirements for substrate processing and subcellular localization pptx
... Coomassie stained and major bands were excised and eluted in order to perform N-terminal sequencing Tandem affinity puri cation and mass spectrometric analysis of USP7 N-terminal and C-terminal TAP-tagged ... N-terminal region motivated us to investigate post-translational modifications on tandem affinity puri cation (TAP)-tagged USP7 purified from mammalian cells LC-MS ⁄ MS analysis revealed the presence ... 208–560 and 208–1102 were amplified with primers containing restriction sites for HindIII and XbaI as 5¢- and 3¢-overhangs, respectively The PCR products obtained were ligated into pCR2. 1-TOPO and...
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Báo cáo khoa học: Human recombinant prolidase from eukaryotic and prokaryotic sources Expression, purification, characterization and long-term stability studies pptx
... prolidase deficiency patients: identi cation of the first PEPD small duplication and of a mutation generating symptomatic and asymptomatic outcome within the same family J Med Genet, in press Fernandez-Espla ... ampli cation was carried out at 94 °C for min, 35 cycles at 94 °C for 30 s, 65 °C for 30 s, and 68 °C for min, and a final extension step at 72 °C for 10 The amplified product was gel-purified using ... synthesis, puri cation and biochemical characterization of human recombinant prolidase from eukaryotic and prokaryotic hosts and compare the enzyme with the endogenous prolidase of human fibroblasts...
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Báo cáo khoa học: Purification, microsequencing and cloning of spinach ATP-dependent phosphofructokinase link sequence and function for the plant enzyme pdf
... sequences from other sources Table Puri cation of spinach ATP-PFK Puri cation step Total activity (mU) Total protein (mg) Specific activity (mUÆmg)1) Puri cation (fold) Crude extract DEAE-Sepharose ... 309-fold purified ATP-PFK, but was accompanied by the loss of 93% of total activity (Table 1) Results and Discussion Puri cation and microsequencing of spinach PFK The present puri cation protocol ... collected and assayed for ATP-PFK activity The gel filtration mass standard (Bio-Rad) was eluted under the same conditions In-gel tryptic digestion, peptide sequencing and protein identi cation Purified...
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Báo cáo khoa học: Purification, characterization and molecular cloning of tyrosinase from the cephalopod mollusk, Illex argentinus docx
... literature [43] The Km and Vmax values for different substrates were obtained from the Hanes–Woolf equation Ó FEBS 2003 Extraction of RNA and first strand cDNA preparation All DNA and RNA manipulations ... ethanol-precipitated, and redissolved in 50 lL of water First strand cDNAs for 5¢- and 3¢-RACE were prepared from the ink sac poly(A)+ RNA using a SMART RACE cDNA Ampli cation kit (Clontech, ... °C, s), annealing (68 °C, 10 s) and elongation (72 °C, min) The amplified products were subcloned into pT7 Blue following puri cation by agarose gel electrophoresis and subjected to DNA sequencing...
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Báo cáo khoa học: Identification, characterization and activation mechanism of a tyrosine kinase of Bacillus anthracis docx
... Glu120, Glu121 (NEED motif) and Glu212, the catalytic Cys167, and the positively charged Arg125, Arg176 and Arg207 (Figs S1B and S4) The phosphorylation sites Tyr155 and Tyr210 are also conserved ... (RU) 300 –0.4 –0.6 0.0 300 a protein kinase and its substrate is transient and of low affinity [35,36] The moderate to low value of the binding affinity and kinetic parameters suggests that binding ... purified McsB and mutant proteins was checked as previously described [37] In brief, 500 ng of the purified McsB and the same amount of McsA was incubated with 10 lCi of [32P]ATP[cP] in a final...
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Báo cáo khoa học: Identification, cloning and characterization of two thioredoxin h isoforms, HvTrxh1 and HvTrxh2, from the barley seed proteome pot
... resulting Vmax and Km values for HvTrxh1 were 2.5 nmolặs)1ặnmol)1 and 13 lM Again, HvTrxh2 and TrxTa resembled each other with Vmax of 2.0 and 1.5 nmolặs)1ặnmol)1 and Km of 44 lM and 37 lM, respectively ... both strands and found to be as expected from the identied TC sequences, and conrmed that the cloning junctions were correct Accession numbers for HvTrxh1 and HvTrxh2 sequences are AY245454 and AY245455, ... were isolated from pCR-h1 and pCR-h2 by digestion with Nde1 and BamHI and ligated into the pET11a expression vector linearized with Nde1 and BamHI, to give pETHvTrxh1 and pETHvTrxh2, respectively...
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Báo cáo Y học: Purification, characterization and subunits identification of the diol dehydratase of Lactobacillus collinoides pot
... the void volume The six fractions Table Puri cation of diol dehydratase of L collinoides Puri cation step Specific Protein Activity activity Yield Puri cation (mg) (U) (UÆmg)1) (%) factor Crude ... ampicillin and 25 lgÆmL)1 kanamycin Puri cation procedures Cellular lysis The protocol for the puri cation of the diol dehydratase of L collinoides was adapted from that of Schutz and Radler ... nucleotide sequence The three bands at 63, 28 and 22 kDa were identified as PduC, PduD and PduE, respectively Regarding the fourth band at 59 kDa (Protein 2), we first supposed that it could represent...
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Báo cáo khoa học: Brassinin oxidase, a fungal detoxifying enzyme to overcome a plant defense – purification, characterization and inhibition potx
... was purified by monitoring BO activity using brassinin as substrate Table indicates the degree of puri cation and yield obtained for each step This puri cation protocol involved four steps: first ... percentage of initial activity and puri cation factors are calculated on the basis of specific activities (lmolÆmin)1 = U) Yield Puri cation step Protein (mg) Activity (mU) Specific activity (mUÆmg)1) Crude ... determined by SDS ⁄ PAGE Fig Detoxi cation of the phytoalexin brassinin by the ‘blackleg’ fungi L maculans (L m.) and L biglobosa (L b.) Table Enzyme yields and puri cation factors for BO Recoveries...
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Báo cáo khoa học: Identification, structure and differential expression of novel pleurocidins clustered on the genome of the winter flounder, Pseudopleuronectes americanus (Walbaum) ppt
... PCR-amplified bands corresponding to pleurocidins WF1, WF2, WF3 and WF4 [5] Positively hybridizing clones were picked and replated until 100% purity was achieved and mapped using BamHI, SstI, XhoI and ... brain, muscle and skin (20–50 mg tissue) of an adult winter flounder, and from pooled samples of 20 whole larvae (hatch and d.p.h.), 10 whole larvae (15, 20, 25 30 and 36 d.p.h.) and five whole juveniles ... not assayed The ampli cation conditions were: at 94 °C; 32 cycles of 30 s at 94 °C, 30 s at 52 °C, 90 s at 72 °C; and at 72 °C Ampli cation of b-actin mRNA was performed to confirm the steady-state...
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Báo cáo khoa học: Purification, characterization and biosynthesis of parabutoxin 3, a component of Parabuthus transvaalicus venom pptx
... to different ion channels and receptors could be performed and a structural model for this novel toxin has been proposed MATERIALS AND METHODS Venom collection and puri cation P transvaalicus scorpions ... constants son and soff for blockade and recovery, current traces were repeated every s before, during and after rPBTx3 application The timecourses of blockade and recovery were fitted to monoexponential ... (s) for the control and the toxin curves are, respectively, 7.5 and 9.6 for Kv1.2, and 9.3 and 8.5 for Kv1.3 channels In all cases, there was no significant shift of V1/2 547 nM and 492 nM, respectively...
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Báo cáo sinh học: " Research Article Audio Signal Processing Using Time-Frequency Approaches: Coding, Classification, Fingerprinting, and Watermarking" pptx
... TFD quanti cation in the next section, and then we explain our proposed TFD quanti cation method 2.3 TFD-Based Quanti cation There have been some attempts in literature to TF quanti cation by ... country, and 25 pop were correctly classified with 100% classi cation accuracy Two out of 21 jazz and out of 34 folk signals were misclassified with a correct classi cation accuracy of 90.5% and 94.1%, ... coupled to statistical classi cation methods 4.1 Music Classi cation In this section, we present a content-based audio retrieval application employing audio classi cation and explain the generic steps...
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Báo cáo khoa học: "Stratification, Blinding and Placebo Effect in a Randomized, Double Blind Placebo-controlled Clinical Trial of Gold Bead Implantation in Dogs with Hip Dysplasia" doc
... of strati cation factor in therapy studies of canine hip dysplasia with a limited number (50-200) of participating dogs A significant placebo effect must be expected, and blinding and randomization ... patients were randomized to receive either gold bead implantation or placebo once in ratio 1:1 by using block randomization with a fixed blocksize of four An error occurred during classi cation of ... signs and medical history using a standardized questionnaire Thereafter, all the dogs were thoroughly examined with special attention to the gait and movement, the lumbar spine and muscles and...
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Báo cáo sinh học: "Changes in muscle cell cation regulation and meat quality traits are associated with genetic selection for high body weight and meat yield in broiler chickens" ppt
... of cations other than calcium may also contribute to the aetiology of skeletal muscle damage, myopathy and ion channel dysfunction, and that the consequent disturbances in cation transport and ... h at 4°C and then both breast muscles (m pectoralis) and both whole thighs (bone and muscles) were cut from the carcass and evaluated for muscle colour, lightness (L*), redness (a*) and yellowness ... occurred between categories and initial and ultimate pH caused by a larger decline in B than L and T lines Initial and final pH were lower (p < 0.01) in B compared with L and T lines (sed within...
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A study on the kinetics of the reaction between chlorpromazine cation radical and pyrogallol
... dyes and colors It is also used as a developer in photography and holography; a mordant for dying wool; a chemical reagent for antimony and bismuth; and as an active reducer for gold, silver and ... of the reaction and concentration of the reactants: Equation ν = k cAmcBn Where cA and cB are concentration of the reactants, A, and B, in a reaction with stoichiometry m and n, and k is the reaction ... reversion, and other reactions causing reduction of shelf life and nutritive value of the food product Table 1-1 shows some of active oxygen and related species Table 1-1 Active oxygen and related...
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Bài thuyết trình về: "Alkali and Halotolerant Catalase from Halomonas sp. SK1: Overexpression in Escherichia coli, Purification, Characterization, and Genetic Modification"
... KHẢO Phucharoen, K., Hoshino, K., Takenaka, Y., and Shinozawa, T., Puri cation, characterization, and gene sequencing of a catalase from an alkali- and halo-tolerant bacterium, Halomonas sp SK1 ... gấp cuộn,… (Yujin E Kim,∗ Mark S Hipp,∗ Andreas Bracher, Manajit Hayer-Hartl, and F Ulrich Hartl 2013 Molecular Chaperone Functions in Protein Folding and Proteostasis Annual Review of Biochemistry ... (2002) Molecular Chaperone Functions in Protein Folding and Proteostasis.Yujin E Kim,∗ Mark S Hipp,∗ Andreas Bracher, Manajit Hayer-Hartl, and F Ulrich Hartl Annu Rev Biochem 2013 82:323–55 This...
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Heavy metal cation retention by unconventional sorbents (red muds and fly ashes)
... and Disposal, p 7.24McGraw-Hill, New York Freeman H M (1988) Standard Handbook of Hazardous Waste Treatment and Disposal, p 6.5McGraw-Hill, New York Gerth J and Bruemmer G (1983) Adsorption and ... Sorption and filtration methods for gas and water purification, sorption mechanisms, pp 91-146 NATO Science Committee, Noordhoff-Leyden, Alphen aan den Rijn, The Netherlands Low K S and Lee C ... and Kamiyama H (1992) Sorption and desorption behavior of cobalt60, strontium-85 and cesium-137 in a porous tuff-mechanisms and kinetics J Nucl Sci Technol 29, 1184-1193 Park Y J., Jung K H and...
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Tài liệu CCIE Routing and Switching Exam Certification Guide P2 docx
... structure for communication between applications It establishes, manages, and terminates communication connections called sessions Communication sessions consist of service requests and responses that ... availability, and distributing information services It also provides synchronization between the peer applications that reside on separate systems Examples of application layer specifications are ... Application Layer (OSI Layer 7) The application layer provides the user or operating system access to the network services It interacts with software applications by identifying communication...
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Tài liệu CCIE Routing and Switching Exam Certification Guide P1 pptx
... Certified requires significant investment in education and preparation by each candidate Moreover, a rigorous and mandatory biyearly recerti cation process ensures the commitment is long lasting and ... areas and provides specialist certifications Each area usually includes a certi cation at the associate, professional, and expert level; the associate level certi cation is the lowest level and ... certi cation is the traditional CCIE certi cation that involves routing and LAN/ WAN switching The CCIE candidate must pass a two-hour written quali cation test and a hands-on one-day lab test to become...
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Tài liệu Báo cáo khoa học: Identification of Ewing’s sarcoma protein as a G-quadruplex DNA- and RNA-binding protein ppt
... RGG3 (lanes and 4) and 32P-labeled Htelo (lanes and 2) or rHtelo (lanes and 4) (B) EMSA was performed with RGG3 (lanes and 4) and 32P-labeled rHtelo (lanes and 2) or mut rHtelo (lanes and 4) (C, ... Identi cation of Ewing’s sarcoma protein Fig Affinity of EWS for binding to G-quadruplex DNA (A) EMSA was performed with EWS (lanes and 4) and 32P-labeled ETS-1 (lanes and 4) or ssDNA L (lanes and ... G-quadruplex DNA and RNA Fig S2 CD spectra of DNAs and RNAs Fig S3 Binding affinity of EWS and RBD to Htelo 998 Fig S4 In vitro arginine methylation of RGG3 by PRMT3 Fig S5 Identi cation of significant residues...
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