xbai Thermo Fisher Search Results


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  • 99
    Thermo Fisher pcdna3 1
    Dose-response relationship for the inhibition of human SMCT2-mediated [ 14 C]nicotinate uptake by unlabeled nicotinate, lactate, and butyrate (A) and inhibition of human SMCT2-mediated nicotinate uptake by NSAIDs (B). Human SMCT2 cDNA was expressed in HRPE cells. Cells transfected with <t>pcDNA3.1</t> vector served to determine endogenous transport. Uptake of [ 14 C]nicotinate (30 μM) was measured in Na + -containing buffer either in the presence of increasing concentrations of unlabelled nicotinate, L-lactate, or butyrate (A) or various NSAIDs (200 μM) (B). Uptake measured in cells transfected with vector alone was subtracted from corresponding uptake measured in cells transfected with cDNA to calculate cDNA-specific uptake. Results are expressed as percentage of control uptake (100 %) measured in the absence of inhibitors. The degree of statistical significance for nicotinate uptake measured in the presence of NSAIDs compared to nicotinate uptake measured in the absence of various NSAIDs in hSMCT2 cDNA-transfected cells is indicated by * P
    Pcdna3 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 49697 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher xbai
    Southern blot analysis of the four STS markers- CtR-431, CtR-594., CtR-496, and AFLP-376. Restriction enzymes used are <t>EcoRI,</t> HinDIII and <t>XbaI</t> . M, molecular weight marker; R resistant genotype Punjab Lal; S susceptible genotype Arka Lohit. Hybridization
    Xbai, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 5807 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher pcdna3
    Western blot of microsomal proteins with the anti– InsP 3 R-I antibody. Rat cerebellar microsomes ( cerMS ) and microsomes isolated from HEK-293 cells transfected with InsP 3 <t>R-pcDNA3</t> ( IP3R ) or with expression vector alone ( pcDNA3 ) were analyzed by Western blotting with T 443 antibody as described in materials and methods . For each microsomal preparation, 20 μg total protein was loaded on the gel. The arrow points to the 260-kD position expected for InsP 3 R.
    Pcdna3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 45292 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher ecori
    Sender plasmids expressed in Escherichia coli BL21. (A) The modular Sender vector allows facile cloning of any <t>EcoRI</t> , XbaI -flanked <t>synthase</t> open reading frame (ORF) and co-expression with mCherry (mCh) from a single bicistronic mRNA. The vector is represented as SBOL [ 44 ] glyphs (top) and a scaled circular map (bottom). (B) Optical density (OD) readings for E . coli cultures expressing each of the Sender plasmids. (C) mCherry expression is shown as end-point RFP signal normalized to OD 600 after an 8 hour growth period.
    Ecori, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12201 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher hindiii
    CSR within sequences upstream of the S μ TR are reduced in Msh2 − / − mice. (a) Twofold dilutions of genomic DNAs were assayed by Sμ/Sγ1 EcoRI DC-PCR and EcoRI–BsrGI DC-PCR. Samples were normalized using nAChR DC-PCR. (b) CSR sites in wild-type or Msh2 −/− mice measured relative to total wild-type CSR (set to 100%). Mean values and SEM of at least four independent experiments are presented. (c) CSR events in Msh2 −/− mice measured by EcoRI, <t>HindIII,</t> and XbaI real-time DC-PCR and compared with wild-type mice. Mean values and SEM are from six independent experiments with two Msh2 −/− mice and seven independent experiments with four wild-type mice.
    Hindiii, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bamhi
    Confirmation of Self-Ligated pP15A, kana R by Digestion Lane 2: digestion of pP15A, kana R vector with <t>BamHI</t> ; lane 3, <t>xbaI;</t> Lane 4, double digestion with BamHI-XbaI; Lane 5, undigested pP15A, kana R vector; Lanes 1 and 6, GeneRuler DNA ladder mix (Thermo Scientific, Waltham, MA, USA).
    Bamhi, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 13718 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher lipofectamine
    Mutating His 29 , His 125 , His 133 and His 158 abolishes GPI-PLD activity His 29 , His 34 , His 56 , His 81 , His 88 , His 98 , His 125 , His 133 , His 158 or His 219 of mouse GPI-PLD was individually mutated to an asparagine residue as described in the Materials and methods section. COS-I cells were transiently transfected with wild-type (WT) or each mutant (5 μg of plasmid DNA) using <t>Lipofectamine™</t> (20 μg). After 24 h, GPI-PLD catalytic activity was determined in the medium and cell lysate ( A ) or GPI-PLD mass in the medium by Western blotting ( B ) as described in the Materials and methods section. Absolute GPI-PLD activity in the medium and cell lysates from cells transfected with wild-type GPI-PLD was 140±118 ( n =6) and 43±20 ( n =6) c.p.m.·h −1 ·(mg of protein) −1 respectively. Results in ( A ) are from three independent experiments performed in triplicate. Results in ( B ) are representative of three experiments.
    Lipofectamine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 72640 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher kpni
    Mutating His 29 , His 125 , His 133 and His 158 abolishes GPI-PLD activity His 29 , His 34 , His 56 , His 81 , His 88 , His 98 , His 125 , His 133 , His 158 or His 219 of mouse GPI-PLD was individually mutated to an asparagine residue as described in the Materials and methods section. COS-I cells were transiently transfected with wild-type (WT) or each mutant (5 μg of plasmid DNA) using <t>Lipofectamine™</t> (20 μg). After 24 h, GPI-PLD catalytic activity was determined in the medium and cell lysate ( A ) or GPI-PLD mass in the medium by Western blotting ( B ) as described in the Materials and methods section. Absolute GPI-PLD activity in the medium and cell lysates from cells transfected with wild-type GPI-PLD was 140±118 ( n =6) and 43±20 ( n =6) c.p.m.·h −1 ·(mg of protein) −1 respectively. Results in ( A ) are from three independent experiments performed in triplicate. Results in ( B ) are representative of three experiments.
    Kpni, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4148 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher xhoi
    Example of a single BIBAC fingerprint generated with five restriction enzymes (the HaeIII fragment ends were not labeled) and four fluorescent-ddNTPs of the SNaPshot Multiplex Ready Reaction Mix on the ABI 3100 DNA analyzer. The blue peaks show the <t>BamHI</t> fragments labeled with ddGTP-dR110; the green peaks show the HindIII fragments labeled with ddATP-dR6G; the black peaks show the XbaI fragments labeled with ddCTP–dTAMRA; the red peaks show the <t>XhoI</t> fragments labeled with ddTTP–dROX; and the orange peaks show the internal size standard LIZ-500.
    Xhoi, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8200 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher t4 dna ligase
    Example of a single BIBAC fingerprint generated with five restriction enzymes (the HaeIII fragment ends were not labeled) and four fluorescent-ddNTPs of the SNaPshot Multiplex Ready Reaction Mix on the ABI 3100 DNA analyzer. The blue peaks show the <t>BamHI</t> fragments labeled with ddGTP-dR110; the green peaks show the HindIII fragments labeled with ddATP-dR6G; the black peaks show the XbaI fragments labeled with ddCTP–dTAMRA; the red peaks show the <t>XhoI</t> fragments labeled with ddTTP–dROX; and the orange peaks show the internal size standard LIZ-500.
    T4 Dna Ligase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 25057 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher pcdna3 1 vector
    Fibulin-1 overexpression decreases apoptosis in various cancer cell lines and bioinformatic analysis of Fibulin-1. (A) Twenty-four hours after transfection with the control <t>(pcDNA3.1)</t> or FBLN1 plasmid (pcDNA3.1-FBLN1C), MHCC97L and SMCC-7721 cells were deprived of serum for 72 h. (B) Twenty-four hours after transfection with FBLN1 siRNA1 or FBLN1 siRNA2, MHCC97L cells were transfected with the control (pcDNA 3.1) or FBLN1 plasmid (pcDNA3.1-FBLN1C) for the next 24 h. Then, the MHCC97L cells were deprived of serum for 48 h. Apoptosis was analyzed using DAPI staining. ** p
    Pcdna3 1 Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 15615 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pcr2 1 topo
    Fibulin-1 overexpression decreases apoptosis in various cancer cell lines and bioinformatic analysis of Fibulin-1. (A) Twenty-four hours after transfection with the control <t>(pcDNA3.1)</t> or FBLN1 plasmid (pcDNA3.1-FBLN1C), MHCC97L and SMCC-7721 cells were deprived of serum for 72 h. (B) Twenty-four hours after transfection with FBLN1 siRNA1 or FBLN1 siRNA2, MHCC97L cells were transfected with the control (pcDNA 3.1) or FBLN1 plasmid (pcDNA3.1-FBLN1C) for the next 24 h. Then, the MHCC97L cells were deprived of serum for 48 h. Apoptosis was analyzed using DAPI staining. ** p
    Pcr2 1 Topo, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 11675 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher megascript t7 kit
    Fibulin-1 overexpression decreases apoptosis in various cancer cell lines and bioinformatic analysis of Fibulin-1. (A) Twenty-four hours after transfection with the control <t>(pcDNA3.1)</t> or FBLN1 plasmid (pcDNA3.1-FBLN1C), MHCC97L and SMCC-7721 cells were deprived of serum for 72 h. (B) Twenty-four hours after transfection with FBLN1 siRNA1 or FBLN1 siRNA2, MHCC97L cells were transfected with the control (pcDNA 3.1) or FBLN1 plasmid (pcDNA3.1-FBLN1C) for the next 24 h. Then, the MHCC97L cells were deprived of serum for 48 h. Apoptosis was analyzed using DAPI staining. ** p
    Megascript T7 Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8073 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher pcr2 1
    Fibulin-1 overexpression decreases apoptosis in various cancer cell lines and bioinformatic analysis of Fibulin-1. (A) Twenty-four hours after transfection with the control <t>(pcDNA3.1)</t> or FBLN1 plasmid (pcDNA3.1-FBLN1C), MHCC97L and SMCC-7721 cells were deprived of serum for 72 h. (B) Twenty-four hours after transfection with FBLN1 siRNA1 or FBLN1 siRNA2, MHCC97L cells were transfected with the control (pcDNA 3.1) or FBLN1 plasmid (pcDNA3.1-FBLN1C) for the next 24 h. Then, the MHCC97L cells were deprived of serum for 48 h. Apoptosis was analyzed using DAPI staining. ** p
    Pcr2 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 12340 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher t7 rna polymerase
    Relationship between known copy numbers of <t>T7</t> RNA (GI and GII) or viral RNA from stool extract (GIV) and the detection threshold. T7 RNA transcripts were made for representative GI and GII strains, and a fecal specimen was used for GIV. (A) Serial dilution
    T7 Rna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12287 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher mmessage mmachine kit
    Relationship between known copy numbers of <t>T7</t> RNA (GI and GII) or viral RNA from stool extract (GIV) and the detection threshold. T7 RNA transcripts were made for representative GI and GII strains, and a fecal specimen was used for GIV. (A) Serial dilution
    Mmessage Mmachine Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 11886 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Dose-response relationship for the inhibition of human SMCT2-mediated [ 14 C]nicotinate uptake by unlabeled nicotinate, lactate, and butyrate (A) and inhibition of human SMCT2-mediated nicotinate uptake by NSAIDs (B). Human SMCT2 cDNA was expressed in HRPE cells. Cells transfected with pcDNA3.1 vector served to determine endogenous transport. Uptake of [ 14 C]nicotinate (30 μM) was measured in Na + -containing buffer either in the presence of increasing concentrations of unlabelled nicotinate, L-lactate, or butyrate (A) or various NSAIDs (200 μM) (B). Uptake measured in cells transfected with vector alone was subtracted from corresponding uptake measured in cells transfected with cDNA to calculate cDNA-specific uptake. Results are expressed as percentage of control uptake (100 %) measured in the absence of inhibitors. The degree of statistical significance for nicotinate uptake measured in the presence of NSAIDs compared to nicotinate uptake measured in the absence of various NSAIDs in hSMCT2 cDNA-transfected cells is indicated by * P

    Journal: Biochimica et biophysica acta

    Article Title: Cloning and Functional Characterization of Human SMCT2 (SLC5A12) and Expression Pattern of the Transporter in Kidney

    doi: 10.1016/j.bbamem.2007.06.031

    Figure Lengend Snippet: Dose-response relationship for the inhibition of human SMCT2-mediated [ 14 C]nicotinate uptake by unlabeled nicotinate, lactate, and butyrate (A) and inhibition of human SMCT2-mediated nicotinate uptake by NSAIDs (B). Human SMCT2 cDNA was expressed in HRPE cells. Cells transfected with pcDNA3.1 vector served to determine endogenous transport. Uptake of [ 14 C]nicotinate (30 μM) was measured in Na + -containing buffer either in the presence of increasing concentrations of unlabelled nicotinate, L-lactate, or butyrate (A) or various NSAIDs (200 μM) (B). Uptake measured in cells transfected with vector alone was subtracted from corresponding uptake measured in cells transfected with cDNA to calculate cDNA-specific uptake. Results are expressed as percentage of control uptake (100 %) measured in the absence of inhibitors. The degree of statistical significance for nicotinate uptake measured in the presence of NSAIDs compared to nicotinate uptake measured in the absence of various NSAIDs in hSMCT2 cDNA-transfected cells is indicated by * P

    Article Snippet: The ~1.8 kb-long amplification product obtained was cloned into pcDNA3.1 (Invitrogen) and pGH19 (kindly provided by Dr. Peter S. Aronson, Yale University) vectors such that the sense transcription of the cloned insert was under the control of T7 promoter.

    Techniques: Inhibition, Transfection, Plasmid Preparation

    Functional expression of human SMCT2 (SLC5A12) in HRPE cells. HRPE cells were transfected with either pcDNA3.1 vector alone (open bars) or human SMCT2 cDNA (closed bars). ( A, B ) Uptake of [ 14 C]lactate (500 μM), [ 14 C]pyruvate (500 μM), and [ 14 C]nicotinate (30 μM) was measured in the presence of Na + . ( C ) Uptake of [ 14 C]nicotinate was measured either in control buffer (25 mM Hepes/Tris, pH 7.5, 5.4 mM KCl, 1.8 mM CaCl 2 , 0.8 mM MgSO 4 , 5 mM glucose, and 140 mM NaCl) or in buffer in which NaCl was replaced with 140 mM of KCl or LiCl. Where indicated, uptake measured in cDNA-transfected cells was significantly different from the corresponding uptake measured in vector-transfected cells (* P

    Journal: Biochimica et biophysica acta

    Article Title: Cloning and Functional Characterization of Human SMCT2 (SLC5A12) and Expression Pattern of the Transporter in Kidney

    doi: 10.1016/j.bbamem.2007.06.031

    Figure Lengend Snippet: Functional expression of human SMCT2 (SLC5A12) in HRPE cells. HRPE cells were transfected with either pcDNA3.1 vector alone (open bars) or human SMCT2 cDNA (closed bars). ( A, B ) Uptake of [ 14 C]lactate (500 μM), [ 14 C]pyruvate (500 μM), and [ 14 C]nicotinate (30 μM) was measured in the presence of Na + . ( C ) Uptake of [ 14 C]nicotinate was measured either in control buffer (25 mM Hepes/Tris, pH 7.5, 5.4 mM KCl, 1.8 mM CaCl 2 , 0.8 mM MgSO 4 , 5 mM glucose, and 140 mM NaCl) or in buffer in which NaCl was replaced with 140 mM of KCl or LiCl. Where indicated, uptake measured in cDNA-transfected cells was significantly different from the corresponding uptake measured in vector-transfected cells (* P

    Article Snippet: The ~1.8 kb-long amplification product obtained was cloned into pcDNA3.1 (Invitrogen) and pGH19 (kindly provided by Dr. Peter S. Aronson, Yale University) vectors such that the sense transcription of the cloned insert was under the control of T7 promoter.

    Techniques: Functional Assay, Expressing, Transfection, Plasmid Preparation

    Southern blot analysis of the four STS markers- CtR-431, CtR-594., CtR-496, and AFLP-376. Restriction enzymes used are EcoRI, HinDIII and XbaI . M, molecular weight marker; R resistant genotype Punjab Lal; S susceptible genotype Arka Lohit. Hybridization

    Journal: 3 Biotech

    Article Title: Sequence-tagged site-based diagnostic markers linked to a novel anthracnose resistance gene RCt1 in chili pepper (Capsicum annuum L.)

    doi: 10.1007/s13205-018-1552-0

    Figure Lengend Snippet: Southern blot analysis of the four STS markers- CtR-431, CtR-594., CtR-496, and AFLP-376. Restriction enzymes used are EcoRI, HinDIII and XbaI . M, molecular weight marker; R resistant genotype Punjab Lal; S susceptible genotype Arka Lohit. Hybridization

    Article Snippet: Three different restriction endonucleases- EcoRI, HinDIII and XbaI (Thermo Fischer Scientific, USA) were used to digest the genomic DNA from the resistant line ‘Punjab Lal’ and the susceptible line ‘Arka Lohit’.

    Techniques: Southern Blot, Molecular Weight, Marker, Hybridization

    Southern hybridization analysis of Ancylostoma caninum genomic DNA to a probe specific for the bandit transposon. The genomic DNAs were cleaved with endonucleases Xba I (lane 1) and Xho I (lane 2). Molecular size standards in kilobase pairs (kb) are shown at the left.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: The bandit, a New DNA Transposon from a Hookworm--Possible Horizontal Genetic Transfer between Host and Parasite

    doi: 10.1371/journal.pntd.0000035

    Figure Lengend Snippet: Southern hybridization analysis of Ancylostoma caninum genomic DNA to a probe specific for the bandit transposon. The genomic DNAs were cleaved with endonucleases Xba I (lane 1) and Xho I (lane 2). Molecular size standards in kilobase pairs (kb) are shown at the left.

    Article Snippet: Briefly, gDNA was digested with the endonuclease Hin d III and Xba I (Fermentas, Sweden) and size separated through 0.8% agarose gel.

    Techniques: Hybridization

    Stable transfection confirms that PAG sequences reduce transcription and steady-state levels of mRNAs from genes in the vicinity. ( A ) Schematic diagram of a bicistronic construct based on the two procyclin genes in the EP/PAG1 locus. Fragments derived from PAG1 were inserted into the unique XbaI site (X) in the forward or reverse orientation. ( B ) Northern blots were hybridized sequentially with probes corresponding to the open reading frames of CAT and BLE, then normalized with α-tubulin. ( C ) Nuclear run-on analysis of BLE and CAT in selected clones. Other single-stranded probes are as described in the legend to Figure 5 . The MARP signals minus background levels were set as 1 for each clone.

    Journal: Nucleic Acids Research

    Article Title: Bidirectional silencing of RNA polymerase I transcription by a strand switch region in Trypanosoma brucei

    doi: 10.1093/nar/gkp513

    Figure Lengend Snippet: Stable transfection confirms that PAG sequences reduce transcription and steady-state levels of mRNAs from genes in the vicinity. ( A ) Schematic diagram of a bicistronic construct based on the two procyclin genes in the EP/PAG1 locus. Fragments derived from PAG1 were inserted into the unique XbaI site (X) in the forward or reverse orientation. ( B ) Northern blots were hybridized sequentially with probes corresponding to the open reading frames of CAT and BLE, then normalized with α-tubulin. ( C ) Nuclear run-on analysis of BLE and CAT in selected clones. Other single-stranded probes are as described in the legend to Figure 5 . The MARP signals minus background levels were set as 1 for each clone.

    Article Snippet: The insert was excised with XhoI (cuts within the vector backbone 5′ to the PAG1 insert) and XbaI and cloned into the XhoI and XbaI sites of pG-mcs-CAT/EP2. pG-1240rev-CAT/EP2 (1240R ): the first 1240 bp of PAG1 were amplified by PCR from pG-1240-CAT/EP2 with the primers PAG1down3 and 1240up3 and cloned into the XhoI and XbaI sites of pG-mcs-CAT/EP2. pG-1240-neo-CAT/EP2 (1240F -neo): the same strategy was used as for pG-640-neo-CAT/EP2, however the neo fragment was cloned into XbaI site of pG-1240-CAT/EP2. pG-PAG1end-CAT/EP2 (ORFF ): to obtain the fragment with the PAG1 sequence from its internal HindIII site at 1087 to position 2458, a PCR was performed with the template pBS-711-HN ( ) and the primers M13r (Invitrogen) and PAG1up2.

    Techniques: Stable Transfection, Construct, Derivative Assay, Northern Blot, Clone Assay

    Colony-PCR and digestion products visualized on 1.2% agarose gels. A: Agarose gel electrophoresis of colony-PCR product. Lane 1: 1968 bp PCR product of fusion fragments, M: 100 bp-plus DNA size marker; B: BamHI and Xba XbaI digestion products of recombinant

    Journal: Reports of Biochemistry & Molecular Biology

    Article Title: Construction of a Novel DNA Vaccine Candidate Encoding an HspX-PPE44-EsxV Fusion Antigen of Mycobacterium tuberculosis

    doi:

    Figure Lengend Snippet: Colony-PCR and digestion products visualized on 1.2% agarose gels. A: Agarose gel electrophoresis of colony-PCR product. Lane 1: 1968 bp PCR product of fusion fragments, M: 100 bp-plus DNA size marker; B: BamHI and Xba XbaI digestion products of recombinant

    Article Snippet: Then the generated HspX-PPE44-EsxV, along with vector, was digested with BamHI and XbaI restriction enzymes and ligated into pcDNA3.1 (+) (Invitrogen, USA) in BamHI and XbaI restriction sites at the 5´and 3´ ends ( ).

    Techniques: Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker, Recombinant

    Agarose gels of recombinant vector. a: digestion of pGH/HspX-PPE44-EsxV by BamHI and XbaI . lane 1: pGH (2940 bp) and HspX-PPE4-EsxV (1968 bp); lane; M: 1kb DNA size marker. B: Ligated pcDNA3.1 (+)/hspX-ppe44-esxV. The band is Recombinant pcDNA3.1 (+)

    Journal: Reports of Biochemistry & Molecular Biology

    Article Title: Construction of a Novel DNA Vaccine Candidate Encoding an HspX-PPE44-EsxV Fusion Antigen of Mycobacterium tuberculosis

    doi:

    Figure Lengend Snippet: Agarose gels of recombinant vector. a: digestion of pGH/HspX-PPE44-EsxV by BamHI and XbaI . lane 1: pGH (2940 bp) and HspX-PPE4-EsxV (1968 bp); lane; M: 1kb DNA size marker. B: Ligated pcDNA3.1 (+)/hspX-ppe44-esxV. The band is Recombinant pcDNA3.1 (+)

    Article Snippet: Then the generated HspX-PPE44-EsxV, along with vector, was digested with BamHI and XbaI restriction enzymes and ligated into pcDNA3.1 (+) (Invitrogen, USA) in BamHI and XbaI restriction sites at the 5´and 3´ ends ( ).

    Techniques: Recombinant, Plasmid Preparation, Marker

    Schematic map of the pcDNA3.1 (+)/HspX-PPE44-EsxV-His plasmid. The fusion segment consisting of hspX, linker, ppe44, linker, esxV sequences, and a 6-polyhistidine-tag was designed between the BamHI and XbaI restriction sites of pcDNA3.1 (+) downstream

    Journal: Reports of Biochemistry & Molecular Biology

    Article Title: Construction of a Novel DNA Vaccine Candidate Encoding an HspX-PPE44-EsxV Fusion Antigen of Mycobacterium tuberculosis

    doi:

    Figure Lengend Snippet: Schematic map of the pcDNA3.1 (+)/HspX-PPE44-EsxV-His plasmid. The fusion segment consisting of hspX, linker, ppe44, linker, esxV sequences, and a 6-polyhistidine-tag was designed between the BamHI and XbaI restriction sites of pcDNA3.1 (+) downstream

    Article Snippet: Then the generated HspX-PPE44-EsxV, along with vector, was digested with BamHI and XbaI restriction enzymes and ligated into pcDNA3.1 (+) (Invitrogen, USA) in BamHI and XbaI restriction sites at the 5´and 3´ ends ( ).

    Techniques: Plasmid Preparation

    Western blot of microsomal proteins with the anti– InsP 3 R-I antibody. Rat cerebellar microsomes ( cerMS ) and microsomes isolated from HEK-293 cells transfected with InsP 3 R-pcDNA3 ( IP3R ) or with expression vector alone ( pcDNA3 ) were analyzed by Western blotting with T 443 antibody as described in materials and methods . For each microsomal preparation, 20 μg total protein was loaded on the gel. The arrow points to the 260-kD position expected for InsP 3 R.

    Journal: The Journal of General Physiology

    Article Title: Single-Channel Properties of Inositol (1,4,5)-Trisphosphate Receptor Heterologously Expressed in HEK-293 Cells

    doi:

    Figure Lengend Snippet: Western blot of microsomal proteins with the anti– InsP 3 R-I antibody. Rat cerebellar microsomes ( cerMS ) and microsomes isolated from HEK-293 cells transfected with InsP 3 R-pcDNA3 ( IP3R ) or with expression vector alone ( pcDNA3 ) were analyzed by Western blotting with T 443 antibody as described in materials and methods . For each microsomal preparation, 20 μg total protein was loaded on the gel. The arrow points to the 260-kD position expected for InsP 3 R.

    Article Snippet: To increase efficiency of InsP3 R-I expression in HEK-293 cells, InsP3 R-I coding sequence was subcloned into pcDNA3 (Invitrogen Corp., San Diego, CA) expression vector and 5′ untranslated region of the original pCMVI-9 clone was substituted with the Kozak consensus sequence ( ).

    Techniques: Western Blot, Isolation, Transfection, Expressing, Plasmid Preparation

    Specific [ 3 H]InsP 3 binding sites are expressed in InsP 3 R-pcDNA3–transfected HEK-293 cells. ( A ) Density of specific [ 3 H]InsP 3 binding sites in microsomal preparations isolated from InsP 3 R-pcDNA3–transfected HEK-293 cells ( IP3R , n = 6), from the untransfected cells ( HEK293 ), from the cells transfected with the expression vector alone ( pcDNA3 ), and from the rat cerebellar microsomes ( cerMS , n = 10). Microsomal preparation and [ 3 H]InsP 3 binding assay were performed as described in materials and methods . The data shown are mean ± SEM. ( B ) Scatchard analysis of specific [ 3 H]InsP 3 binding sites present in microsomes isolated from InsP 3 R-pcDNA3–transfected HEK-293 cells. Fit to the data ( line ) yields K d = 60 nM InsP 3 and B max = 12 pmol/mg.

    Journal: The Journal of General Physiology

    Article Title: Single-Channel Properties of Inositol (1,4,5)-Trisphosphate Receptor Heterologously Expressed in HEK-293 Cells

    doi:

    Figure Lengend Snippet: Specific [ 3 H]InsP 3 binding sites are expressed in InsP 3 R-pcDNA3–transfected HEK-293 cells. ( A ) Density of specific [ 3 H]InsP 3 binding sites in microsomal preparations isolated from InsP 3 R-pcDNA3–transfected HEK-293 cells ( IP3R , n = 6), from the untransfected cells ( HEK293 ), from the cells transfected with the expression vector alone ( pcDNA3 ), and from the rat cerebellar microsomes ( cerMS , n = 10). Microsomal preparation and [ 3 H]InsP 3 binding assay were performed as described in materials and methods . The data shown are mean ± SEM. ( B ) Scatchard analysis of specific [ 3 H]InsP 3 binding sites present in microsomes isolated from InsP 3 R-pcDNA3–transfected HEK-293 cells. Fit to the data ( line ) yields K d = 60 nM InsP 3 and B max = 12 pmol/mg.

    Article Snippet: To increase efficiency of InsP3 R-I expression in HEK-293 cells, InsP3 R-I coding sequence was subcloned into pcDNA3 (Invitrogen Corp., San Diego, CA) expression vector and 5′ untranslated region of the original pCMVI-9 clone was substituted with the Kozak consensus sequence ( ).

    Techniques: Binding Assay, Transfection, Isolation, Expressing, Plasmid Preparation

    Single channel records of recombinant InsP 3 R and native rat cerebellar InsP 3 R in planar lipid bilayers. Representative single channel currents through recombinant rat InsP 3 R ( A ) and native rat cerebellar InsP 3 R ( B ). Control traces were obtained in the presence of 0.2 μM Ca 2+ and 1 mM Na 2 ATP. Addition of 2 μM InsP 3 on the cis (cytosolic) side evoked openings of recombinant InsP 3 R-I and native rat cerebellar InsP 3 R. Current traces at the expanded time scale are shown on the bottom. Similar results were obtained with 7 independent microsomal preparations from InsP 3 R-I-transfected HEK-293 cells and 10 rat cerebellar microsomal preparations. InsP 3 R-gated channels were not observed in similar reconstitution experiments with microsomes from untransfected HEK-293 cells or from HEK-293 transfected with pcDNA3 vector alone.

    Journal: The Journal of General Physiology

    Article Title: Single-Channel Properties of Inositol (1,4,5)-Trisphosphate Receptor Heterologously Expressed in HEK-293 Cells

    doi:

    Figure Lengend Snippet: Single channel records of recombinant InsP 3 R and native rat cerebellar InsP 3 R in planar lipid bilayers. Representative single channel currents through recombinant rat InsP 3 R ( A ) and native rat cerebellar InsP 3 R ( B ). Control traces were obtained in the presence of 0.2 μM Ca 2+ and 1 mM Na 2 ATP. Addition of 2 μM InsP 3 on the cis (cytosolic) side evoked openings of recombinant InsP 3 R-I and native rat cerebellar InsP 3 R. Current traces at the expanded time scale are shown on the bottom. Similar results were obtained with 7 independent microsomal preparations from InsP 3 R-I-transfected HEK-293 cells and 10 rat cerebellar microsomal preparations. InsP 3 R-gated channels were not observed in similar reconstitution experiments with microsomes from untransfected HEK-293 cells or from HEK-293 transfected with pcDNA3 vector alone.

    Article Snippet: To increase efficiency of InsP3 R-I expression in HEK-293 cells, InsP3 R-I coding sequence was subcloned into pcDNA3 (Invitrogen Corp., San Diego, CA) expression vector and 5′ untranslated region of the original pCMVI-9 clone was substituted with the Kozak consensus sequence ( ).

    Techniques: Recombinant, Transfection, Plasmid Preparation

    Immunocytochemical staining of transfected HEK-293 cells with the anti–InsP 3 R-I antibody. HEK-293 cells were transfected with InsP 3 R-pcDNA3 clone ( A ) or pcDNA3 vector alone ( B ). Expressed InsP 3 R-I receptors were detected by immunocytochemical staining using T 443 antibody and rhodamine-conjugated anti–rabbit IgG. Immunostaining and fluorescent imaging was performed as described in materials and methods using 20× objective. Fluorescent signal was monitored on rhodamine channel.

    Journal: The Journal of General Physiology

    Article Title: Single-Channel Properties of Inositol (1,4,5)-Trisphosphate Receptor Heterologously Expressed in HEK-293 Cells

    doi:

    Figure Lengend Snippet: Immunocytochemical staining of transfected HEK-293 cells with the anti–InsP 3 R-I antibody. HEK-293 cells were transfected with InsP 3 R-pcDNA3 clone ( A ) or pcDNA3 vector alone ( B ). Expressed InsP 3 R-I receptors were detected by immunocytochemical staining using T 443 antibody and rhodamine-conjugated anti–rabbit IgG. Immunostaining and fluorescent imaging was performed as described in materials and methods using 20× objective. Fluorescent signal was monitored on rhodamine channel.

    Article Snippet: To increase efficiency of InsP3 R-I expression in HEK-293 cells, InsP3 R-I coding sequence was subcloned into pcDNA3 (Invitrogen Corp., San Diego, CA) expression vector and 5′ untranslated region of the original pCMVI-9 clone was substituted with the Kozak consensus sequence ( ).

    Techniques: Staining, Transfection, Plasmid Preparation, Immunostaining, Imaging

    Sender plasmids expressed in Escherichia coli BL21. (A) The modular Sender vector allows facile cloning of any EcoRI , XbaI -flanked synthase open reading frame (ORF) and co-expression with mCherry (mCh) from a single bicistronic mRNA. The vector is represented as SBOL [ 44 ] glyphs (top) and a scaled circular map (bottom). (B) Optical density (OD) readings for E . coli cultures expressing each of the Sender plasmids. (C) mCherry expression is shown as end-point RFP signal normalized to OD 600 after an 8 hour growth period.

    Journal: PLoS ONE

    Article Title: Characterization of diverse homoserine lactone synthases in Escherichia coli

    doi: 10.1371/journal.pone.0202294

    Figure Lengend Snippet: Sender plasmids expressed in Escherichia coli BL21. (A) The modular Sender vector allows facile cloning of any EcoRI , XbaI -flanked synthase open reading frame (ORF) and co-expression with mCherry (mCh) from a single bicistronic mRNA. The vector is represented as SBOL [ 44 ] glyphs (top) and a scaled circular map (bottom). (B) Optical density (OD) readings for E . coli cultures expressing each of the Sender plasmids. (C) mCherry expression is shown as end-point RFP signal normalized to OD 600 after an 8 hour growth period.

    Article Snippet: The Modular Sender Vector and each HSL synthase oligo were cut with EcoRI and XbaI (Thermo Fisher Scientific) and ligated using T4 ligase (New England Biolabs).

    Techniques: Plasmid Preparation, Clone Assay, Expressing

    CSR within sequences upstream of the S μ TR are reduced in Msh2 − / − mice. (a) Twofold dilutions of genomic DNAs were assayed by Sμ/Sγ1 EcoRI DC-PCR and EcoRI–BsrGI DC-PCR. Samples were normalized using nAChR DC-PCR. (b) CSR sites in wild-type or Msh2 −/− mice measured relative to total wild-type CSR (set to 100%). Mean values and SEM of at least four independent experiments are presented. (c) CSR events in Msh2 −/− mice measured by EcoRI, HindIII, and XbaI real-time DC-PCR and compared with wild-type mice. Mean values and SEM are from six independent experiments with two Msh2 −/− mice and seven independent experiments with four wild-type mice.

    Journal: The Journal of Experimental Medicine

    Article Title: Shifts in targeting of class switch recombination sites in mice that lack ? switch region tandem repeats or Msh2

    doi: 10.1084/jem.20042491

    Figure Lengend Snippet: CSR within sequences upstream of the S μ TR are reduced in Msh2 − / − mice. (a) Twofold dilutions of genomic DNAs were assayed by Sμ/Sγ1 EcoRI DC-PCR and EcoRI–BsrGI DC-PCR. Samples were normalized using nAChR DC-PCR. (b) CSR sites in wild-type or Msh2 −/− mice measured relative to total wild-type CSR (set to 100%). Mean values and SEM of at least four independent experiments are presented. (c) CSR events in Msh2 −/− mice measured by EcoRI, HindIII, and XbaI real-time DC-PCR and compared with wild-type mice. Mean values and SEM are from six independent experiments with two Msh2 −/− mice and seven independent experiments with four wild-type mice.

    Article Snippet: For switch events located downstream of the SμTR, genomic DNAs were digested with EcoRI, HindIII, or XbaI, and circularized for real-time PCR using fluorescent SYBR green dye (Applied Biosystems).

    Techniques: Mouse Assay, Polymerase Chain Reaction

    S μ TR − / − mice have increased levels of CSR events targeted to downstream sequences. (a) DC-PCR measuring CSR junction sites downstream of the SμTR region. HindIII and XbaI sites downstream of the SμTR region were used in DC-PCR assays to measure CSR junctions in sequences downstream of each site. Total CSR sites were determined by EcoRI DC-PCR (E, EcoRI; H1 and H2, HindIII; X, XbaI). Primer pairs used for the different DC-PCR assays are indicated by small arrows of different types. (b) Wild-type and SμTR −/− CSR events measured by EcoRI, HindIII, and XbaI real-time DC-PCR. Copy numbers for CSR events were derived from pSμ/Sγ3 standard curves. Rag-1 PCR normalized CSR events relative to Rag-1 copy number. Mean values and SEM are from seven independent experiments with four mice per group. (c) Percentage of SμTR −/− switch events downstream of the H2 relative to events downstream of H1 (set to 100%). Mean values and SEM are from three independent experiments per group.

    Journal: The Journal of Experimental Medicine

    Article Title: Shifts in targeting of class switch recombination sites in mice that lack ? switch region tandem repeats or Msh2

    doi: 10.1084/jem.20042491

    Figure Lengend Snippet: S μ TR − / − mice have increased levels of CSR events targeted to downstream sequences. (a) DC-PCR measuring CSR junction sites downstream of the SμTR region. HindIII and XbaI sites downstream of the SμTR region were used in DC-PCR assays to measure CSR junctions in sequences downstream of each site. Total CSR sites were determined by EcoRI DC-PCR (E, EcoRI; H1 and H2, HindIII; X, XbaI). Primer pairs used for the different DC-PCR assays are indicated by small arrows of different types. (b) Wild-type and SμTR −/− CSR events measured by EcoRI, HindIII, and XbaI real-time DC-PCR. Copy numbers for CSR events were derived from pSμ/Sγ3 standard curves. Rag-1 PCR normalized CSR events relative to Rag-1 copy number. Mean values and SEM are from seven independent experiments with four mice per group. (c) Percentage of SμTR −/− switch events downstream of the H2 relative to events downstream of H1 (set to 100%). Mean values and SEM are from three independent experiments per group.

    Article Snippet: For switch events located downstream of the SμTR, genomic DNAs were digested with EcoRI, HindIII, or XbaI, and circularized for real-time PCR using fluorescent SYBR green dye (Applied Biosystems).

    Techniques: Mouse Assay, Polymerase Chain Reaction, Derivative Assay

    Confirmation of Self-Ligated pP15A, kana R by Digestion Lane 2: digestion of pP15A, kana R vector with BamHI ; lane 3, xbaI; Lane 4, double digestion with BamHI-XbaI; Lane 5, undigested pP15A, kana R vector; Lanes 1 and 6, GeneRuler DNA ladder mix (Thermo Scientific, Waltham, MA, USA).

    Journal: Jundishapur Journal of Microbiology

    Article Title: Zinc Finger Nuclease: A New Approach to Overcome Beta-Lactam Antibiotic Resistance

    doi: 10.5812/jjm.29384

    Figure Lengend Snippet: Confirmation of Self-Ligated pP15A, kana R by Digestion Lane 2: digestion of pP15A, kana R vector with BamHI ; lane 3, xbaI; Lane 4, double digestion with BamHI-XbaI; Lane 5, undigested pP15A, kana R vector; Lanes 1 and 6, GeneRuler DNA ladder mix (Thermo Scientific, Waltham, MA, USA).

    Article Snippet: The purified plasmid was confirmed by single and double-digestion reactions by BamHI and XbaI restriction enzymes (Thermo Scientific, Waltham, MA, USA) and after confirmation was used to clone and express the left and right ZFN arrays.

    Techniques: Plasmid Preparation

    Mutating His 29 , His 125 , His 133 and His 158 abolishes GPI-PLD activity His 29 , His 34 , His 56 , His 81 , His 88 , His 98 , His 125 , His 133 , His 158 or His 219 of mouse GPI-PLD was individually mutated to an asparagine residue as described in the Materials and methods section. COS-I cells were transiently transfected with wild-type (WT) or each mutant (5 μg of plasmid DNA) using Lipofectamine™ (20 μg). After 24 h, GPI-PLD catalytic activity was determined in the medium and cell lysate ( A ) or GPI-PLD mass in the medium by Western blotting ( B ) as described in the Materials and methods section. Absolute GPI-PLD activity in the medium and cell lysates from cells transfected with wild-type GPI-PLD was 140±118 ( n =6) and 43±20 ( n =6) c.p.m.·h −1 ·(mg of protein) −1 respectively. Results in ( A ) are from three independent experiments performed in triplicate. Results in ( B ) are representative of three experiments.

    Journal: Biochemical Journal

    Article Title: Mutating His29, His125, His133 or His158 abolishes glycosylphosphatidylinositol-specific phospholipase D catalytic activity

    doi: 10.1042/BJ20050656

    Figure Lengend Snippet: Mutating His 29 , His 125 , His 133 and His 158 abolishes GPI-PLD activity His 29 , His 34 , His 56 , His 81 , His 88 , His 98 , His 125 , His 133 , His 158 or His 219 of mouse GPI-PLD was individually mutated to an asparagine residue as described in the Materials and methods section. COS-I cells were transiently transfected with wild-type (WT) or each mutant (5 μg of plasmid DNA) using Lipofectamine™ (20 μg). After 24 h, GPI-PLD catalytic activity was determined in the medium and cell lysate ( A ) or GPI-PLD mass in the medium by Western blotting ( B ) as described in the Materials and methods section. Absolute GPI-PLD activity in the medium and cell lysates from cells transfected with wild-type GPI-PLD was 140±118 ( n =6) and 43±20 ( n =6) c.p.m.·h −1 ·(mg of protein) −1 respectively. Results in ( A ) are from three independent experiments performed in triplicate. Results in ( B ) are representative of three experiments.

    Article Snippet: COS-I cells (A.T.C.C., Rockville, MD, U.S.A.) (60 mm dishes) were transiently transfected with wild-type or each mutant (5 μg of plasmid cDNA) using 20 μg of Lipofectamine™ (Invitrogen) in Opti-MEM® I (Reduced Serum Medium) according to the manufacturer's methods.

    Techniques: Activity Assay, Transfection, Mutagenesis, Plasmid Preparation, Western Blot

    Example of a single BIBAC fingerprint generated with five restriction enzymes (the HaeIII fragment ends were not labeled) and four fluorescent-ddNTPs of the SNaPshot Multiplex Ready Reaction Mix on the ABI 3100 DNA analyzer. The blue peaks show the BamHI fragments labeled with ddGTP-dR110; the green peaks show the HindIII fragments labeled with ddATP-dR6G; the black peaks show the XbaI fragments labeled with ddCTP–dTAMRA; the red peaks show the XhoI fragments labeled with ddTTP–dROX; and the orange peaks show the internal size standard LIZ-500.

    Journal: Nucleic Acids Research

    Article Title: Genome physical mapping from large-insert clones by fingerprint analysis with capillary electrophoresis: a robust physical map of Penicillium chrysogenum

    doi: 10.1093/nar/gni037

    Figure Lengend Snippet: Example of a single BIBAC fingerprint generated with five restriction enzymes (the HaeIII fragment ends were not labeled) and four fluorescent-ddNTPs of the SNaPshot Multiplex Ready Reaction Mix on the ABI 3100 DNA analyzer. The blue peaks show the BamHI fragments labeled with ddGTP-dR110; the green peaks show the HindIII fragments labeled with ddATP-dR6G; the black peaks show the XbaI fragments labeled with ddCTP–dTAMRA; the red peaks show the XhoI fragments labeled with ddTTP–dROX; and the orange peaks show the internal size standard LIZ-500.

    Article Snippet: The DNA was digested and end-labeled using a five-enzyme, four-color labeling kit consisting of (i) four 6 bp restriction endonucleases, BamHI, Hind III, XbaI and XhoI, producing four different 3′ recessed ends to be labeled, (ii) one 4 bp restriction endonuclease, HaeIII, producing blunted ends and (iii) the SNaPshot Multiplex Ready Reaction Mix (Applied Biosystems).

    Techniques: Generated, Labeling, Multiplex Assay

    Fibulin-1 overexpression decreases apoptosis in various cancer cell lines and bioinformatic analysis of Fibulin-1. (A) Twenty-four hours after transfection with the control (pcDNA3.1) or FBLN1 plasmid (pcDNA3.1-FBLN1C), MHCC97L and SMCC-7721 cells were deprived of serum for 72 h. (B) Twenty-four hours after transfection with FBLN1 siRNA1 or FBLN1 siRNA2, MHCC97L cells were transfected with the control (pcDNA 3.1) or FBLN1 plasmid (pcDNA3.1-FBLN1C) for the next 24 h. Then, the MHCC97L cells were deprived of serum for 48 h. Apoptosis was analyzed using DAPI staining. ** p

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Increased Expression of Fibulin-1 Is Associated With Hepatocellular Carcinoma Progression by Regulating the Notch Signaling Pathway

    doi: 10.3389/fcell.2020.00478

    Figure Lengend Snippet: Fibulin-1 overexpression decreases apoptosis in various cancer cell lines and bioinformatic analysis of Fibulin-1. (A) Twenty-four hours after transfection with the control (pcDNA3.1) or FBLN1 plasmid (pcDNA3.1-FBLN1C), MHCC97L and SMCC-7721 cells were deprived of serum for 72 h. (B) Twenty-four hours after transfection with FBLN1 siRNA1 or FBLN1 siRNA2, MHCC97L cells were transfected with the control (pcDNA 3.1) or FBLN1 plasmid (pcDNA3.1-FBLN1C) for the next 24 h. Then, the MHCC97L cells were deprived of serum for 48 h. Apoptosis was analyzed using DAPI staining. ** p

    Article Snippet: The coding sequence of the FBLN1 gene was PCR-amplified and inserted into the Eco RI/XbaI sites of the pcDNA3.1 vector (Life Technologies), as previously described ( ).

    Techniques: Over Expression, Transfection, Plasmid Preparation, Staining

    Western blot assays of nuclear extracts from astrocytic cell cultures (PDA) transfected with pCDNA3 (lane 1), pCDNA3NF-1X (NF-1X; lane 2), or pET-c-Jun (c-Jun; lane 3) and subsequently infected with JCV. Equal amounts of extract were resolved on each lane of a 4 to 12% gradient gel, transferred to a PVDF membrane, and then probed with anti-Vp-1, anti-NF-1X, anti-c-Jun, and anti-β-actin. β-Actin detection served as a control for equivalent total protein loading.

    Journal: Journal of Virology

    Article Title: Interactions between c-Jun, Nuclear Factor 1, and JC Virus Promoter Sequences: Implications for Viral Tropism ▿

    doi: 10.1128/JVI.01355-06

    Figure Lengend Snippet: Western blot assays of nuclear extracts from astrocytic cell cultures (PDA) transfected with pCDNA3 (lane 1), pCDNA3NF-1X (NF-1X; lane 2), or pET-c-Jun (c-Jun; lane 3) and subsequently infected with JCV. Equal amounts of extract were resolved on each lane of a 4 to 12% gradient gel, transferred to a PVDF membrane, and then probed with anti-Vp-1, anti-NF-1X, anti-c-Jun, and anti-β-actin. β-Actin detection served as a control for equivalent total protein loading.

    Article Snippet: The pcDNA3NF-1X plasmid was subcloned by forced cloning of the hemagglutinin (HA)-tagged NF-1X2 gene from pCHAmNF-1X2 (described below) into the multicloning site (between the NotI and XbaI restriction sites) of the pcDNA3 vector (Invitrogen). pCHAmNF1-X2 was a generous gift from Richard Gronostajski (The State University of New York at Buffalo) which contains the cDNA coding region of murine NF-1X (with a single amino acid difference, murine NF-1X is > 99.7% homologous to human NF-1X).

    Techniques: Western Blot, Transfection, Positron Emission Tomography, Infection

    Relationship between known copy numbers of T7 RNA (GI and GII) or viral RNA from stool extract (GIV) and the detection threshold. T7 RNA transcripts were made for representative GI and GII strains, and a fecal specimen was used for GIV. (A) Serial dilution

    Journal: Journal of Clinical Microbiology

    Article Title: Use of TaqMan Real-Time Reverse Transcription-PCR for Rapid Detection, Quantification, and Typing of Norovirus

    doi: 10.1128/JCM.44.4.1405-1412.2006

    Figure Lengend Snippet: Relationship between known copy numbers of T7 RNA (GI and GII) or viral RNA from stool extract (GIV) and the detection threshold. T7 RNA transcripts were made for representative GI and GII strains, and a fecal specimen was used for GIV. (A) Serial dilution

    Article Snippet: After the cloned plasmid DNA was purified by use of the QIAfilter plasmid Mega kit (QIAGEN Inc., Valencia, CA), the DNA insert was cut out with the restriction enzymes, gel purified, and used as the template for in vitro transcription with T7 RNA polymerase with the MEGAscript kit (Ambion Inc., Austin, TX).

    Techniques: Serial Dilution