taq polymerase Takara Search Results


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  • 99
    TaKaRa takara taq polymerase
    Construction of RV M33 virus full-length cDNA clone. The numbers on the viral genome scale refer to the distance from the 5′ end in kilobases. Six DNA fragments were amplified by proofreading <t>TaKaRa</t> <t>Taq</t> DNA polymerase with the requisite primers as described in Materials and Methods. The amplified DNA fragments were ligated into a full-length cDNA representing the whole viral genome by using the restriction sites indicated above the genome. The full-length cDNA was cut with Eco RI and Hin dIII and inserted into a modified pBR322 plasmid that had been cut with the same enzymes to obtain the full-length cDNA clone pBRM33.
    Takara Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 2349 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    TaKaRa l a taq polymerase
    PCR of subdivided genomic sequences. Three regions of human genomic DNA (GenBank accession nos AC006454 , AC093734 and X91835 , with 5103, 5193 and 12 114 bp, respectively) were subdivided into nine 567, 577 and 1346 bp PCR sites, respectively. PCR was performed for each subdivided site using primer sets (20 bp each) corresponding to the terminal sequence of each site using the <t>Taq</t> DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; <t>Takara-bio).</t> PCR was carried out in the absence and in the presence of Tth RecA protein and ATP. The products were electrophoresed and stained with ethidium bromide. ( a ) A diagrammatic representation of the subdivided region (5103 bp in GenBank accession no AC006454 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( b ) A diagrammatic representation of the subdivided region (5193 bp in GenBank accession no AC093734 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( c ) A diagrammatic representation of the subdivided region (12 114 bp in GenBank accession no X91835 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). Throughout (a–c), nine subdivided sites for each region are indicated as a-1 to a-9, b-1 to b-9 and c-1 to c-9. Nucleotide (nt) numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.
    L A Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 87/100, based on 202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa takara taq
    PCR of subdivided genomic sequences. Three regions of human genomic DNA (GenBank accession nos AC006454 , AC093734 and X91835 , with 5103, 5193 and 12 114 bp, respectively) were subdivided into nine 567, 577 and 1346 bp PCR sites, respectively. PCR was performed for each subdivided site using primer sets (20 bp each) corresponding to the terminal sequence of each site using the <t>Taq</t> DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; <t>Takara-bio).</t> PCR was carried out in the absence and in the presence of Tth RecA protein and ATP. The products were electrophoresed and stained with ethidium bromide. ( a ) A diagrammatic representation of the subdivided region (5103 bp in GenBank accession no AC006454 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( b ) A diagrammatic representation of the subdivided region (5193 bp in GenBank accession no AC093734 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( c ) A diagrammatic representation of the subdivided region (12 114 bp in GenBank accession no X91835 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). Throughout (a–c), nine subdivided sites for each region are indicated as a-1 to a-9, b-1 to b-9 and c-1 to c-9. Nucleotide (nt) numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.
    Takara Taq, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 602 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    TaKaRa gxl taq polymerase
    PCR of subdivided genomic sequences. Three regions of human genomic DNA (GenBank accession nos AC006454 , AC093734 and X91835 , with 5103, 5193 and 12 114 bp, respectively) were subdivided into nine 567, 577 and 1346 bp PCR sites, respectively. PCR was performed for each subdivided site using primer sets (20 bp each) corresponding to the terminal sequence of each site using the <t>Taq</t> DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; <t>Takara-bio).</t> PCR was carried out in the absence and in the presence of Tth RecA protein and ATP. The products were electrophoresed and stained with ethidium bromide. ( a ) A diagrammatic representation of the subdivided region (5103 bp in GenBank accession no AC006454 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( b ) A diagrammatic representation of the subdivided region (5193 bp in GenBank accession no AC093734 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( c ) A diagrammatic representation of the subdivided region (12 114 bp in GenBank accession no X91835 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). Throughout (a–c), nine subdivided sites for each region are indicated as a-1 to a-9, b-1 to b-9 and c-1 to c-9. Nucleotide (nt) numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.
    Gxl Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa start taq polymerase
    PCR of subdivided genomic sequences. Three regions of human genomic DNA (GenBank accession nos AC006454 , AC093734 and X91835 , with 5103, 5193 and 12 114 bp, respectively) were subdivided into nine 567, 577 and 1346 bp PCR sites, respectively. PCR was performed for each subdivided site using primer sets (20 bp each) corresponding to the terminal sequence of each site using the <t>Taq</t> DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; <t>Takara-bio).</t> PCR was carried out in the absence and in the presence of Tth RecA protein and ATP. The products were electrophoresed and stained with ethidium bromide. ( a ) A diagrammatic representation of the subdivided region (5103 bp in GenBank accession no AC006454 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( b ) A diagrammatic representation of the subdivided region (5193 bp in GenBank accession no AC093734 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( c ) A diagrammatic representation of the subdivided region (12 114 bp in GenBank accession no X91835 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). Throughout (a–c), nine subdivided sites for each region are indicated as a-1 to a-9, b-1 to b-9 and c-1 to c-9. Nucleotide (nt) numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.
    Start Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TaKaRa taq polymerase buffer
    PCR of subdivided genomic sequences. Three regions of human genomic DNA (GenBank accession nos AC006454 , AC093734 and X91835 , with 5103, 5193 and 12 114 bp, respectively) were subdivided into nine 567, 577 and 1346 bp PCR sites, respectively. PCR was performed for each subdivided site using primer sets (20 bp each) corresponding to the terminal sequence of each site using the <t>Taq</t> DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; <t>Takara-bio).</t> PCR was carried out in the absence and in the presence of Tth RecA protein and ATP. The products were electrophoresed and stained with ethidium bromide. ( a ) A diagrammatic representation of the subdivided region (5103 bp in GenBank accession no AC006454 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( b ) A diagrammatic representation of the subdivided region (5193 bp in GenBank accession no AC093734 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( c ) A diagrammatic representation of the subdivided region (12 114 bp in GenBank accession no X91835 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). Throughout (a–c), nine subdivided sites for each region are indicated as a-1 to a-9, b-1 to b-9 and c-1 to c-9. Nucleotide (nt) numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.
    Taq Polymerase Buffer, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 89 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa takara ex taq polymerase
    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with <t>Taq</t> I. Lane M: 100 bp <t>DNA</t> ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.
    Takara Ex Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1239 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa premix taq dna polymerase
    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with <t>Taq</t> I. Lane M: 100 bp <t>DNA</t> ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.
    Premix Taq Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 255 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa takara la taq polymerase
    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with <t>Taq</t> I. Lane M: 100 bp <t>DNA</t> ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.
    Takara La Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa z taq polymerase
    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with <t>Taq</t> I. Lane M: 100 bp <t>DNA</t> ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.
    Z Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    TaKaRa hotstar taq polymerase
    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with <t>Taq</t> I. Lane M: 100 bp <t>DNA</t> ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.
    Hotstar Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher takara taq polymerase
    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with <t>Taq</t> I. Lane M: 100 bp <t>DNA</t> ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.
    Takara Taq Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa takara la taq
    Impaired µ–γ1 switch recombination of DC3-5Rα –/– B cells in response to IL-5 plus CS/2. Splenic B cells (2·5 × 10 6 in a 5-ml culture) were cultured with CS/2 (0·5 µg/ml) or CS/2 (0·5 µg/ml) plus IL-5 (100 U/ml). On day 2, cells were harvested, the cell number was adjusted to 2·5 × 10 6 /5 ml and the cells were re-cultured with relevant stimuli for 24 hr. On day 3, total DNA were prepared from live cells and the DNA samples (50 and 100 ng) were amplified using <t>5′Sγ1</t> and 3′Sµ primers and LA- <t>Taq</t> polymerase and hybridized with 5′Sγ1 probes.
    Takara La Taq, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 2811 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Construction of RV M33 virus full-length cDNA clone. The numbers on the viral genome scale refer to the distance from the 5′ end in kilobases. Six DNA fragments were amplified by proofreading TaKaRa Taq DNA polymerase with the requisite primers as described in Materials and Methods. The amplified DNA fragments were ligated into a full-length cDNA representing the whole viral genome by using the restriction sites indicated above the genome. The full-length cDNA was cut with Eco RI and Hin dIII and inserted into a modified pBR322 plasmid that had been cut with the same enzymes to obtain the full-length cDNA clone pBRM33.

    Journal: Journal of Virology

    Article Title: Mutational Analysis, Using a Full-Length Rubella Virus cDNA Clone, of Rubella Virus E1 Transmembrane and Cytoplasmic Domains Required for Virus Release

    doi:

    Figure Lengend Snippet: Construction of RV M33 virus full-length cDNA clone. The numbers on the viral genome scale refer to the distance from the 5′ end in kilobases. Six DNA fragments were amplified by proofreading TaKaRa Taq DNA polymerase with the requisite primers as described in Materials and Methods. The amplified DNA fragments were ligated into a full-length cDNA representing the whole viral genome by using the restriction sites indicated above the genome. The full-length cDNA was cut with Eco RI and Hin dIII and inserted into a modified pBR322 plasmid that had been cut with the same enzymes to obtain the full-length cDNA clone pBRM33.

    Article Snippet: PCR amplifications were performed in separate reactions containing cDNA, 1 pmol of primers, 0.4 mM deoxynucleoside triphosphates, 10% dimethyl sulfoxide, and proofreading TaKaRa Taq polymerase (Takara Shuzo Co., Ltd.) in a buffer provided by the manufacturer.

    Techniques: Amplification, Modification, Plasmid Preparation

    (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic DNA. PCR was performed on the isolated genomic DNA using taq DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for

    Journal: Neuroscience letters

    Article Title: Involvement of GluR2 and GluR3 subunit C-termini in the trigeminal spinal subnucleus caudalis and C1-C2 neurons in trigeminal neuropathic pain

    doi: 10.1016/j.neulet.2010.12.060

    Figure Lengend Snippet: (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic DNA. PCR was performed on the isolated genomic DNA using taq DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for

    Article Snippet: PCR was performed on the isolated genomic DNA using taq DNA polymerase (TaKaRa Ex Taq™, Takara, Otsu, Japan) and primer sets (primer #21–23 for GluR2 delta7: #21, 5′-ACA GAG GAA GGT AGT GGA AGG GAG-3′; #22, 5′-CTT GGT TTG GTT GTT GGT CAT AGC-3′; #23, 5′-CTA GTG AAC CTC TTC GAG GGA C-3′; primer #31–33 for GluR3 delta7: #31, 5′-CCA ATA CTC CAC AGG GGC AAT TTA TC-3′; #32, 5′-CCG TTG ACT GTT TTG AAT CTC ACA CC-3′; #33, 5′-CTA GTG AAC CTC TTC GAG GGA C-3′).

    Techniques: Mutagenesis, Mouse Assay, Polymerase Chain Reaction, Isolation

    Agarose gel electrophoresis of napA gene amplified by PCR. Lane 1: DL2000 DNA marker; lane 2: H pylori napA gene (435 bp).

    Journal:

    Article Title: Detection and evaluation of antibodies against neutrophil-activating protein of Helicobacter pylori in patients with gastric cancer

    doi: 10.3748/wjg.15.2381

    Figure Lengend Snippet: Agarose gel electrophoresis of napA gene amplified by PCR. Lane 1: DL2000 DNA marker; lane 2: H pylori napA gene (435 bp).

    Article Snippet: The extracted genomic DNA was then used as a template for amplification of the NAP coding region using a Taq DNA polymerase PCR kit (Takara, Japan)[ , ].

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

    Agarose gel electrophoresis analysis of PCR product of napA gene from H pylori clinic strains. Lane 1: DL2000 DNA marker; Lanes 2-9: napA gene (435 bp) from eight H pylori clinic strains.

    Journal:

    Article Title: Detection and evaluation of antibodies against neutrophil-activating protein of Helicobacter pylori in patients with gastric cancer

    doi: 10.3748/wjg.15.2381

    Figure Lengend Snippet: Agarose gel electrophoresis analysis of PCR product of napA gene from H pylori clinic strains. Lane 1: DL2000 DNA marker; Lanes 2-9: napA gene (435 bp) from eight H pylori clinic strains.

    Article Snippet: The extracted genomic DNA was then used as a template for amplification of the NAP coding region using a Taq DNA polymerase PCR kit (Takara, Japan)[ , ].

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Marker

    DNA fingerprint analysis of specific scFv genes of positive clones . scFv inserts were amplified from individual colonies of all positive clones. Amplification was done with Ex-Taq DNA polymearase enzyme using pCANTAB5-R1 and pCANTAB5-S6 primers. The amplified products were then analyzed on agarose gels. A . scFv encoding inserts of DNA of 40 diversified positive clones B . Only five restriction patterns of scFv encoding inserts of DNA were identified against five different clones (scFv K1 - K5). Molecular weights of DNA ladder markers (kbp) were indicated by M on the left.

    Journal: BMC Biotechnology

    Article Title: An improved phage-display panning method to produce an HM-1 killer toxin anti-idiotypic antibody

    doi: 10.1186/1472-6750-9-99

    Figure Lengend Snippet: DNA fingerprint analysis of specific scFv genes of positive clones . scFv inserts were amplified from individual colonies of all positive clones. Amplification was done with Ex-Taq DNA polymearase enzyme using pCANTAB5-R1 and pCANTAB5-S6 primers. The amplified products were then analyzed on agarose gels. A . scFv encoding inserts of DNA of 40 diversified positive clones B . Only five restriction patterns of scFv encoding inserts of DNA were identified against five different clones (scFv K1 - K5). Molecular weights of DNA ladder markers (kbp) were indicated by M on the left.

    Article Snippet: Plasmid DNAs from anti-idiotypic antibody producing clones were isolated from E. coli HB2151 by alkaline lysis and the scFv gene insert of individual clones was amplified by using PCR with pCANTAB5-R1 primer (5'-CCATGATTACGCCAAGCTTTGGAGCC-3') and pCANTAB5-S6 primer (5'-GTAAATGAATTTTCTGTATGAGG-3') and with Ex-Taq DNA polymerase enzyme (Takara).

    Techniques: Clone Assay, Amplification

    PCR of subdivided genomic sequences. Three regions of human genomic DNA (GenBank accession nos AC006454 , AC093734 and X91835 , with 5103, 5193 and 12 114 bp, respectively) were subdivided into nine 567, 577 and 1346 bp PCR sites, respectively. PCR was performed for each subdivided site using primer sets (20 bp each) corresponding to the terminal sequence of each site using the Taq DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; Takara-bio). PCR was carried out in the absence and in the presence of Tth RecA protein and ATP. The products were electrophoresed and stained with ethidium bromide. ( a ) A diagrammatic representation of the subdivided region (5103 bp in GenBank accession no AC006454 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( b ) A diagrammatic representation of the subdivided region (5193 bp in GenBank accession no AC093734 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( c ) A diagrammatic representation of the subdivided region (12 114 bp in GenBank accession no X91835 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). Throughout (a–c), nine subdivided sites for each region are indicated as a-1 to a-9, b-1 to b-9 and c-1 to c-9. Nucleotide (nt) numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.

    Journal: Nucleic Acids Research

    Article Title: Multiplex PCR: use of heat-stable Thermus thermophilus RecA protein to minimize non-specific PCR products

    doi: 10.1093/nar/gni111

    Figure Lengend Snippet: PCR of subdivided genomic sequences. Three regions of human genomic DNA (GenBank accession nos AC006454 , AC093734 and X91835 , with 5103, 5193 and 12 114 bp, respectively) were subdivided into nine 567, 577 and 1346 bp PCR sites, respectively. PCR was performed for each subdivided site using primer sets (20 bp each) corresponding to the terminal sequence of each site using the Taq DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; Takara-bio). PCR was carried out in the absence and in the presence of Tth RecA protein and ATP. The products were electrophoresed and stained with ethidium bromide. ( a ) A diagrammatic representation of the subdivided region (5103 bp in GenBank accession no AC006454 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( b ) A diagrammatic representation of the subdivided region (5193 bp in GenBank accession no AC093734 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). ( c ) A diagrammatic representation of the subdivided region (12 114 bp in GenBank accession no X91835 ) (upper panel) and the electrophoretic patterns of the PCR products (lower panel). Throughout (a–c), nine subdivided sites for each region are indicated as a-1 to a-9, b-1 to b-9 and c-1 to c-9. Nucleotide (nt) numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.

    Article Snippet: The same results were obtained by the other polymerase systems: LA Taq polymerase (Takara-bio), Tth polymerase (Applied-boisystems); Expand High Fidelity, Expand High FidelityPLUS and Expand Long Template polymerase (Roche-diagnostics); TITANIUM Taq polymerase (Becton-Dickinson-Clontech); and Taq polymerase (Promega).

    Techniques: Polymerase Chain Reaction, Genomic Sequencing, Sequencing, Staining

    PCR with primers carrying mismatched bases. PCR was performed at two human genomic sites with primers (20 bp), one of which (forward primer) carried one, two or three mismatched bases in the middle of the primer, in the absence (left) or presence (right) of Tth RecA protein and ATP using the Taq DNA polymerase ( rTaq DNA polymerase plus ‘hot start’ antibody). ( a ) Upper panel: a diagrammatic representation of the location of the PCR site (20 bp between nt 66 562 and nt 66 581 in GenBank accession no AC006454 ) and of the primers. Lower panel: lanes 1 and 5, PCR products using primers without mismatched bases (primer set a-1); lanes 2 and 6, PCR products using primers (primer set a-2 with one mismatched base at nt 66 566, T to C); lanes 3 and 7, PCR products using primers (primer set a-3 with two mismatched base at nt 66 566 and nt 66 571, both T to C); and lanes 4 and 8, PCR products using primers (primer set a-4 with three mismatched base at nt 66 566 and nt 66 571, T to C and nt 66 576, G to C). The oligonucleotide sequences used for the forward primers (mismatched bases are underlined) are as follows: primer set a-1, 5′-CATGGCACCTGCTCTGAGAC-3′; primer set a-2, 5′-CATGGCACC C GCTCTGAGAC-3′; primer set a-3, 5′-CATGGCACC C GCTC C GAGAC-3′; and primer set a-4, 5′-CATG C CACC C GCTC C GAGAC-3′. ( b ) Upper panel: a diagrammatic representation of the location of the PCR site (20 bp between nt 38 501 and nt 38 520 in GenBank accession no. AC0937734 ) and of the primers. Lower panel: lanes 1 and 5, PCR products using primers without mismatched bases (primer set b-1); lanes 2 and 6, PCR products using primers (primer set b-2 with one mismatched base at nt 38 505, G to A); lanes 3 and 7, PCR products using primers (primer set b-3 with two mismatched base at nt 38 505 and nt 38 510, both G to A); and lanes 4 and 8, PCR products using primers (primer set b-4 with three mismatched base at nt 38 505, nt 38 510 and nt 38 515, all G to A). The oligonucleotide sequences used for the forward primers are as follows: primer set b-1, 5′-ATCTGTGTGGTTCGGCTCTG-3′; primer set b-2, 5′-ATCTGTGTG A TTCGGCTCTG-3′; primer set b-3, 5′-ATCTGTGTG A TTCG A CTCTG-3′; and primer set b-4, 5′-ATCT A TGTG A TTCG A CTCTG-3′. ( c ) Upper panel: a diagrammatic representation of the location of the PCR site (20 bp between nt 63 957 and nt 63 976 in GenBank accession no. AC004975 ) and of the primers. Lower panel: lanes 1 and 5, PCR products using primers without mismatched bases (primer set c-1); lanes 2 and 6, PCR products using primers (primer set c-2 with one mismatched base at nt 63 961, A to T); lanes 3 and 7, PCR products using primers (primer set c-3 with two mismatched base at nt 63 961 and nt 63 966, A to T and C to T); and lanes 4 and 8, PCR products using primers (primer set c-4 with three mismatched base at nt 63 961, nt 63 966 and nt 63 971, A to T, C to T and G to T). The oligonucleotide sequences used for the forward primers are as follows: primer set c-1, 5′-GCAGGCACCAAGAACTACTG-3′; primer set c-2, 5′-GCAGGCACC T AGAACTACTG-3′; primer set c-3, 5′-GCAGGCACC T AGAA T TACTG-3′; and primer set c-4, 5′-GCAG T CACC T AGAA T TACTG-3′. The sequences for the backward primers are 5′-TCACCTCCCAGCCTGGCCCA-3′ for ( a ), 5′-AGGGAGATGTTCTCATAAAT-3′ and 5′-CTGTAAGTGGCAGACATTAC-3′ for ( b ). Nucleotide numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.

    Journal: Nucleic Acids Research

    Article Title: Multiplex PCR: use of heat-stable Thermus thermophilus RecA protein to minimize non-specific PCR products

    doi: 10.1093/nar/gni111

    Figure Lengend Snippet: PCR with primers carrying mismatched bases. PCR was performed at two human genomic sites with primers (20 bp), one of which (forward primer) carried one, two or three mismatched bases in the middle of the primer, in the absence (left) or presence (right) of Tth RecA protein and ATP using the Taq DNA polymerase ( rTaq DNA polymerase plus ‘hot start’ antibody). ( a ) Upper panel: a diagrammatic representation of the location of the PCR site (20 bp between nt 66 562 and nt 66 581 in GenBank accession no AC006454 ) and of the primers. Lower panel: lanes 1 and 5, PCR products using primers without mismatched bases (primer set a-1); lanes 2 and 6, PCR products using primers (primer set a-2 with one mismatched base at nt 66 566, T to C); lanes 3 and 7, PCR products using primers (primer set a-3 with two mismatched base at nt 66 566 and nt 66 571, both T to C); and lanes 4 and 8, PCR products using primers (primer set a-4 with three mismatched base at nt 66 566 and nt 66 571, T to C and nt 66 576, G to C). The oligonucleotide sequences used for the forward primers (mismatched bases are underlined) are as follows: primer set a-1, 5′-CATGGCACCTGCTCTGAGAC-3′; primer set a-2, 5′-CATGGCACC C GCTCTGAGAC-3′; primer set a-3, 5′-CATGGCACC C GCTC C GAGAC-3′; and primer set a-4, 5′-CATG C CACC C GCTC C GAGAC-3′. ( b ) Upper panel: a diagrammatic representation of the location of the PCR site (20 bp between nt 38 501 and nt 38 520 in GenBank accession no. AC0937734 ) and of the primers. Lower panel: lanes 1 and 5, PCR products using primers without mismatched bases (primer set b-1); lanes 2 and 6, PCR products using primers (primer set b-2 with one mismatched base at nt 38 505, G to A); lanes 3 and 7, PCR products using primers (primer set b-3 with two mismatched base at nt 38 505 and nt 38 510, both G to A); and lanes 4 and 8, PCR products using primers (primer set b-4 with three mismatched base at nt 38 505, nt 38 510 and nt 38 515, all G to A). The oligonucleotide sequences used for the forward primers are as follows: primer set b-1, 5′-ATCTGTGTGGTTCGGCTCTG-3′; primer set b-2, 5′-ATCTGTGTG A TTCGGCTCTG-3′; primer set b-3, 5′-ATCTGTGTG A TTCG A CTCTG-3′; and primer set b-4, 5′-ATCT A TGTG A TTCG A CTCTG-3′. ( c ) Upper panel: a diagrammatic representation of the location of the PCR site (20 bp between nt 63 957 and nt 63 976 in GenBank accession no. AC004975 ) and of the primers. Lower panel: lanes 1 and 5, PCR products using primers without mismatched bases (primer set c-1); lanes 2 and 6, PCR products using primers (primer set c-2 with one mismatched base at nt 63 961, A to T); lanes 3 and 7, PCR products using primers (primer set c-3 with two mismatched base at nt 63 961 and nt 63 966, A to T and C to T); and lanes 4 and 8, PCR products using primers (primer set c-4 with three mismatched base at nt 63 961, nt 63 966 and nt 63 971, A to T, C to T and G to T). The oligonucleotide sequences used for the forward primers are as follows: primer set c-1, 5′-GCAGGCACCAAGAACTACTG-3′; primer set c-2, 5′-GCAGGCACC T AGAACTACTG-3′; primer set c-3, 5′-GCAGGCACC T AGAA T TACTG-3′; and primer set c-4, 5′-GCAG T CACC T AGAA T TACTG-3′. The sequences for the backward primers are 5′-TCACCTCCCAGCCTGGCCCA-3′ for ( a ), 5′-AGGGAGATGTTCTCATAAAT-3′ and 5′-CTGTAAGTGGCAGACATTAC-3′ for ( b ). Nucleotide numbers correspond to registries in GenBank. Locations of the specific PCR products are indicated by arrows.

    Article Snippet: The same results were obtained by the other polymerase systems: LA Taq polymerase (Takara-bio), Tth polymerase (Applied-boisystems); Expand High Fidelity, Expand High FidelityPLUS and Expand Long Template polymerase (Roche-diagnostics); TITANIUM Taq polymerase (Becton-Dickinson-Clontech); and Taq polymerase (Promega).

    Techniques: Polymerase Chain Reaction

    Effect of T.thermophilus RecA protein on PCR. PCR with Taq DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; Takara-bio) for several randomly selected sequences (300–650 bp) in human genomic DNA was carried out in the absence or in the presence of the Tth RecA protein. ( a ) Control, PCR under the standard conditions described in Materials and Methods. ( b ) Similar to (a), but with Tth RecA protein (0.4 μg per 25 μl reaction mixture). ( c ) Similar to (a), but with ATP (400 μM). ( d ) Similar to (a), but with Tth RecA protein (0.4 μg per 25 μl reaction mixture) and ATP (300 μM). ( e ) Similar to (a), but with Tth RecA protein (0.4 μg per 25 μl reaction mixture) and ATP-γS (300 μM). The products were electrophoresed and stained with ethidium bromide. Molecular weight markers are indicated on the right-hand side of these panels. The oligonucleotide sequences used for the primers were as follows: 5′-ACAATGGGCTCACTCACCCA-3′ and 5′-CTAAGACCAATGGATAGCTG-3′ for lane 1 (300 bp); 5′-GCTCAGCATGGTGGTGGCAT-3′ and 5′-CCTCATACCTTCCCCCCCAT-3′ for lane 2 (319 bp); 5′-GACTACTCTAGCGACTGTCC-3′ and 5′-GACAGCCACCAGATCCAATC-3′ for lane 3 (360 bp); 5′-AACCTCACAACCTTGGCTGA-3′ and 5′-TTCACAACTTAAGATTTGGC-3′ for lane 4 (400 bp); 5′-AGGCAACTAGGATGGTGTGG-3′ and 5′-CAGGGAGCGTGTCCATAGGG-3′ for lane 5 (450 bp); 5′-CTGCTGAAAGAGATGCGGTG-3′ and 5′-AGGAAAACAGCCCAAGGGAC-3′ for lane 6 (469 bp); and 5′-ACTTTGTTCTGAGCCTCACA-3′ and 5′-GTTGCCCAATCGCCCCTCTC-3′ for lane 7 (650 bp).

    Journal: Nucleic Acids Research

    Article Title: Multiplex PCR: use of heat-stable Thermus thermophilus RecA protein to minimize non-specific PCR products

    doi: 10.1093/nar/gni111

    Figure Lengend Snippet: Effect of T.thermophilus RecA protein on PCR. PCR with Taq DNA polymerase ( ExTaq DNA polymerase plus ‘hot start’ antibody; Takara-bio) for several randomly selected sequences (300–650 bp) in human genomic DNA was carried out in the absence or in the presence of the Tth RecA protein. ( a ) Control, PCR under the standard conditions described in Materials and Methods. ( b ) Similar to (a), but with Tth RecA protein (0.4 μg per 25 μl reaction mixture). ( c ) Similar to (a), but with ATP (400 μM). ( d ) Similar to (a), but with Tth RecA protein (0.4 μg per 25 μl reaction mixture) and ATP (300 μM). ( e ) Similar to (a), but with Tth RecA protein (0.4 μg per 25 μl reaction mixture) and ATP-γS (300 μM). The products were electrophoresed and stained with ethidium bromide. Molecular weight markers are indicated on the right-hand side of these panels. The oligonucleotide sequences used for the primers were as follows: 5′-ACAATGGGCTCACTCACCCA-3′ and 5′-CTAAGACCAATGGATAGCTG-3′ for lane 1 (300 bp); 5′-GCTCAGCATGGTGGTGGCAT-3′ and 5′-CCTCATACCTTCCCCCCCAT-3′ for lane 2 (319 bp); 5′-GACTACTCTAGCGACTGTCC-3′ and 5′-GACAGCCACCAGATCCAATC-3′ for lane 3 (360 bp); 5′-AACCTCACAACCTTGGCTGA-3′ and 5′-TTCACAACTTAAGATTTGGC-3′ for lane 4 (400 bp); 5′-AGGCAACTAGGATGGTGTGG-3′ and 5′-CAGGGAGCGTGTCCATAGGG-3′ for lane 5 (450 bp); 5′-CTGCTGAAAGAGATGCGGTG-3′ and 5′-AGGAAAACAGCCCAAGGGAC-3′ for lane 6 (469 bp); and 5′-ACTTTGTTCTGAGCCTCACA-3′ and 5′-GTTGCCCAATCGCCCCTCTC-3′ for lane 7 (650 bp).

    Article Snippet: The same results were obtained by the other polymerase systems: LA Taq polymerase (Takara-bio), Tth polymerase (Applied-boisystems); Expand High Fidelity, Expand High FidelityPLUS and Expand Long Template polymerase (Roche-diagnostics); TITANIUM Taq polymerase (Becton-Dickinson-Clontech); and Taq polymerase (Promega).

    Techniques: Polymerase Chain Reaction, Staining, Molecular Weight

    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with Taq I. Lane M: 100 bp DNA ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.

    Journal: Journal of Veterinary Science

    Article Title: The 23S rRNA gene PCR-RFLP used for characterization of porcine intestinal spirochete isolates

    doi: 10.4142/jvs.2006.7.3.277

    Figure Lengend Snippet: PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with Taq I. Lane M: 100 bp DNA ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.

    Article Snippet: The PCR conditions consisted of 5 µl (50 ng/µl) of DNA and 1 µl each of primer (50 pM) in a 5 µl of 10× reaction buffer with 5 µl of 25 mM MgCl2 , 5 µl of 10 mM dNTP (each 2.5 mM) and 1 µl of 5 U Ex Taq DNA polymerase (TaKaRa, Japan) to a final volume of 50 µl on a thermal cycler (PTC-100; MJ Research, USA).

    Techniques: Polymerase Chain Reaction, Agarose Gel Electrophoresis

    Impaired µ–γ1 switch recombination of DC3-5Rα –/– B cells in response to IL-5 plus CS/2. Splenic B cells (2·5 × 10 6 in a 5-ml culture) were cultured with CS/2 (0·5 µg/ml) or CS/2 (0·5 µg/ml) plus IL-5 (100 U/ml). On day 2, cells were harvested, the cell number was adjusted to 2·5 × 10 6 /5 ml and the cells were re-cultured with relevant stimuli for 24 hr. On day 3, total DNA were prepared from live cells and the DNA samples (50 and 100 ng) were amplified using 5′Sγ1 and 3′Sµ primers and LA- Taq polymerase and hybridized with 5′Sγ1 probes.

    Journal: Immunology

    Article Title: Functional dissection of the cytoplasmic subregions of the interleukin-5 receptor ? chain in growth and immunoglobulin G1 switch recombination of B cells

    doi: 10.1046/j.1365-2567.2001.01196.x

    Figure Lengend Snippet: Impaired µ–γ1 switch recombination of DC3-5Rα –/– B cells in response to IL-5 plus CS/2. Splenic B cells (2·5 × 10 6 in a 5-ml culture) were cultured with CS/2 (0·5 µg/ml) or CS/2 (0·5 µg/ml) plus IL-5 (100 U/ml). On day 2, cells were harvested, the cell number was adjusted to 2·5 × 10 6 /5 ml and the cells were re-cultured with relevant stimuli for 24 hr. On day 3, total DNA were prepared from live cells and the DNA samples (50 and 100 ng) were amplified using 5′Sγ1 and 3′Sµ primers and LA- Taq polymerase and hybridized with 5′Sγ1 probes.

    Article Snippet: For amplification of γ1–µ recombination products, 200 ng of total DNA from cultured or freshly isolated B cells were subjected to PCR amplification in 50 µl volumes containing LA PCR Buffer II (Takara; Kyoto, Japan), 2·5 m m MgCl2 , 0·4 m m dNTP, 2·5 U LA Taq (Takara), 1 µ m sense-strand 5′Sγ1 primer (5′-CACTCCTGGGTATGGAAACACATCCTAC-3′; nucleotides 262–289 in region 5′ of the Sγ1 repeats; MUSIGHANB) in combination with an anti-sense 3′Sµ primer (5′-AGCCTAACTTATCTGAGCCTAGTTCAAC-3′; nucleotides 1347–1319 in region 3′ of the Sµ repeats; MUSIGCD09).

    Techniques: Cell Culture, Amplification