hotstar taq dna polymerase kit  (Qiagen)


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    Structured Review

    Qiagen hotstar taq dna polymerase kit
    Hotstar Taq Dna Polymerase Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 93/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hotstar taq dna polymerase kit/product/Qiagen
    Average 93 stars, based on 38 article reviews
    Price from $9.99 to $1999.99
    hotstar taq dna polymerase kit - by Bioz Stars, 2020-04
    93/100 stars

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    Related Articles

    Diagnostic Assay:

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: The gene-specific primer (GSP) used for PCR amplification of the target cDNA was the diagnostic forward primer, 5 -GGGCCTAATCACGATGACC-3 , mentioned above. .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Article Title: Glucocorticoids employ the monomeric glucocorticoid receptor to potentiate vitamin D3 and parathyroid hormone–induced osteoclastogenesis
    Article Snippet: Materials Mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), recombinant mouse M-CSF, recombinant extracellular domain of mouse RANKL (Arg72-Asp316; 462-TR), recombinant RANK/TNFRSF11A (RANKL neutralizing soluble RANK), and the ELISA kits for mouse RANKL and mouse OPG were purchased from R & D Systems (Minneapolis, MN, USA). .. HotStar Taq Polymerase Kit, QiaQuick PCR Purification Kit and RNeasy Mini Kit were from Qiagen (Hilden, Germany); DNA-Free and RNAqueous–4PCR Kit were obtained from Ambion (Austin, TX, USA); First-Strand cDNA Synthesis Kit and the PCR Core Kit were from Hoffmann-La Roche (Basel, Switzerland).

    Amplification:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: The gene-specific primer (5′-ACCCAT-GAAGGTAAGAAGCCC-3′) used for PCR amplification of the target cDNA was based on the initially derived 5′ sequence of the N gene. .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing.

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: The abridged universal amplification primer (AUAP) provided in the kit was used as reverse primer. .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing.

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Fluorescence:

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Synthesized:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: Complementary DNA was synthesized from 5 μl of fecal RNA template with an oligo-(dT)-containing adapter primer according to the kit manufacturer's protocol. .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing.

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: Complementary DNA was synthesized from 3 l of fecal RNA template with the oligo(dT)-containing adapter primer provided in the kit, according to the manufacturer's recommended protocol. .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing.

    Chromosome Walking:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing. .. Sequence data for the approximately 2 kb product were obtained by initially using the gene-specific primer described above and a poly-T sequencing primer, followed by primer-walking with newly designed 5′ and 3′ primers derived with the OLIGO 6 primer analysis software (Molecular Biology Insights, Cascade, CO).

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing. .. Sequence data for the 2.5 kb product were obtained by initially using the diagnostic forward primer (GSP) and a poly-T sequencing primer, followed by primer-walking with newly designed 5 and 3 primers derived with the OLIGO 6 software (Molecular Biology Insights, Cascade, CO).

    Purification:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing. .. Sequence data for the approximately 2 kb product were obtained by initially using the gene-specific primer described above and a poly-T sequencing primer, followed by primer-walking with newly designed 5′ and 3′ primers derived with the OLIGO 6 primer analysis software (Molecular Biology Insights, Cascade, CO).

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing. .. Sequence data for the 2.5 kb product were obtained by initially using the diagnostic forward primer (GSP) and a poly-T sequencing primer, followed by primer-walking with newly designed 5 and 3 primers derived with the OLIGO 6 software (Molecular Biology Insights, Cascade, CO).

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Article Title: Glucocorticoids employ the monomeric glucocorticoid receptor to potentiate vitamin D3 and parathyroid hormone–induced osteoclastogenesis
    Article Snippet: .. HotStar Taq Polymerase Kit, QiaQuick PCR Purification Kit and RNeasy Mini Kit were from Qiagen (Hilden, Germany); DNA-Free and RNAqueous–4PCR Kit were obtained from Ambion (Austin, TX, USA); First-Strand cDNA Synthesis Kit and the PCR Core Kit were from Hoffmann-La Roche (Basel, Switzerland). .. Fluorescent-labeled probes (reporter fluorescent dye VIC at the 5′end and quencher fluorescent dye Tamra at the 3′end), TaqMan Universal PCR Master Mix, and the kits for real-time quantitative PCR were from Thermo Fisher Scientific; culture dishes, multiwell plates, and glass chamber slides were from Thermo Fisher Scientific; suspension culture dishes from Corning (Corning, NY, USA); bone slices and CrossLabs for Culture ELISA (CTX) were from Immunodiagnostic Systems (East Boldon, United Kingdom); and the mouse bone marrow stromal cell line ST-2 was from Riken BioResource Research Center Cell Bank (Tsukuba, Japan).

    Real-time Polymerase Chain Reaction:

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Article Title: Glucocorticoids employ the monomeric glucocorticoid receptor to potentiate vitamin D3 and parathyroid hormone–induced osteoclastogenesis
    Article Snippet: HotStar Taq Polymerase Kit, QiaQuick PCR Purification Kit and RNeasy Mini Kit were from Qiagen (Hilden, Germany); DNA-Free and RNAqueous–4PCR Kit were obtained from Ambion (Austin, TX, USA); First-Strand cDNA Synthesis Kit and the PCR Core Kit were from Hoffmann-La Roche (Basel, Switzerland). .. Fluorescent-labeled probes (reporter fluorescent dye VIC at the 5′end and quencher fluorescent dye Tamra at the 3′end), TaqMan Universal PCR Master Mix, and the kits for real-time quantitative PCR were from Thermo Fisher Scientific; culture dishes, multiwell plates, and glass chamber slides were from Thermo Fisher Scientific; suspension culture dishes from Corning (Corning, NY, USA); bone slices and CrossLabs for Culture ELISA (CTX) were from Immunodiagnostic Systems (East Boldon, United Kingdom); and the mouse bone marrow stromal cell line ST-2 was from Riken BioResource Research Center Cell Bank (Tsukuba, Japan).

    Concentration Assay:

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. D3 and indomethacin were dissolved in ethanol; the final concentration of ethanol never exceeded 0.1% and did not by itself affect calcium 45 (45 Ca) release in mouse calvariae.

    Generated:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: Sequencing of the entire N gene A 683-nucleotide sequence (excluding primer sequences) of the ferret coronavirus, which spans the last 334 nucleotides of the M gene, 16 nucleotides of intergenic sequence, and the first 333 nucleotides of the N gene, was obtained by sequencing of amplicons generated with consensus primers. .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing.

    Polymerase Chain Reaction:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing. .. Sequence data for the approximately 2 kb product were obtained by initially using the gene-specific primer described above and a poly-T sequencing primer, followed by primer-walking with newly designed 5′ and 3′ primers derived with the OLIGO 6 primer analysis software (Molecular Biology Insights, Cascade, CO).

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing. .. Sequence data for the 2.5 kb product were obtained by initially using the diagnostic forward primer (GSP) and a poly-T sequencing primer, followed by primer-walking with newly designed 5 and 3 primers derived with the OLIGO 6 software (Molecular Biology Insights, Cascade, CO).

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Article Title: Glucocorticoids employ the monomeric glucocorticoid receptor to potentiate vitamin D3 and parathyroid hormone–induced osteoclastogenesis
    Article Snippet: .. HotStar Taq Polymerase Kit, QiaQuick PCR Purification Kit and RNeasy Mini Kit were from Qiagen (Hilden, Germany); DNA-Free and RNAqueous–4PCR Kit were obtained from Ambion (Austin, TX, USA); First-Strand cDNA Synthesis Kit and the PCR Core Kit were from Hoffmann-La Roche (Basel, Switzerland). .. Fluorescent-labeled probes (reporter fluorescent dye VIC at the 5′end and quencher fluorescent dye Tamra at the 3′end), TaqMan Universal PCR Master Mix, and the kits for real-time quantitative PCR were from Thermo Fisher Scientific; culture dishes, multiwell plates, and glass chamber slides were from Thermo Fisher Scientific; suspension culture dishes from Corning (Corning, NY, USA); bone slices and CrossLabs for Culture ELISA (CTX) were from Immunodiagnostic Systems (East Boldon, United Kingdom); and the mouse bone marrow stromal cell line ST-2 was from Riken BioResource Research Center Cell Bank (Tsukuba, Japan).

    Expressing:

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Gel Extraction:

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing. .. Sequence data for the 2.5 kb product were obtained by initially using the diagnostic forward primer (GSP) and a poly-T sequencing primer, followed by primer-walking with newly designed 5 and 3 primers derived with the OLIGO 6 software (Molecular Biology Insights, Cascade, CO).

    Sequencing:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing. .. Sequence data for the approximately 2 kb product were obtained by initially using the gene-specific primer described above and a poly-T sequencing primer, followed by primer-walking with newly designed 5′ and 3′ primers derived with the OLIGO 6 primer analysis software (Molecular Biology Insights, Cascade, CO).

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing. .. Sequence data for the 2.5 kb product were obtained by initially using the diagnostic forward primer (GSP) and a poly-T sequencing primer, followed by primer-walking with newly designed 5 and 3 primers derived with the OLIGO 6 software (Molecular Biology Insights, Cascade, CO).

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Article Title: Glucocorticoids employ the monomeric glucocorticoid receptor to potentiate vitamin D3 and parathyroid hormone–induced osteoclastogenesis
    Article Snippet: Bovine PTH 1–34 was from Bachem (Bubendorf, Switzerland); α-modification of minimum essential medium (α-MEM), and fetal calf serum were from Thermo Fisher Scientific (Waltham, MA, USA); Thermo Sequenase- II DYEnamic ET Terminator Cycle Sequencing Kit was from Amersham (Little Chalfont, United Kingdom); and oligonucleotide primers were from Thermo Fisher Scientific. .. HotStar Taq Polymerase Kit, QiaQuick PCR Purification Kit and RNeasy Mini Kit were from Qiagen (Hilden, Germany); DNA-Free and RNAqueous–4PCR Kit were obtained from Ambion (Austin, TX, USA); First-Strand cDNA Synthesis Kit and the PCR Core Kit were from Hoffmann-La Roche (Basel, Switzerland).

    Staining:

    Article Title: Glucocorticoids employ the monomeric glucocorticoid receptor to potentiate vitamin D3 and parathyroid hormone–induced osteoclastogenesis
    Article Snippet: The kit for leukocyte acid phosphatase staining, Sigma 104 phosphatase substrate, Trizol, DEX, hydrocortisone, prednisolone, and RU38486 were from MilliporeSigma (Burlington, MA, USA. .. HotStar Taq Polymerase Kit, QiaQuick PCR Purification Kit and RNeasy Mini Kit were from Qiagen (Hilden, Germany); DNA-Free and RNAqueous–4PCR Kit were obtained from Ambion (Austin, TX, USA); First-Strand cDNA Synthesis Kit and the PCR Core Kit were from Hoffmann-La Roche (Basel, Switzerland).

    Recombinant:

    Article Title: Effects on osteoclast and osteoblast activities in cultured mouse calvarial bones by synovial fluids from patients with a loose joint prosthesis and from osteoarthritis patients
    Article Snippet: .. Materials Synthetic bovine parathyroid hormone [PTH-(1–34)] was obtained from Bachem AG (Bubendorf, Switzerland); recombinant human IL-1α, recombinant human IL-1β, recombinant human IL-6, recombinant human soluble IL-6 receptor, recombinant human IL-17, recombinant human TNF-α, mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), antisera neutralising human IL-1α, human IL-1β, human soluble IL-6 receptor, human IL-17, or human TNF-α from R & D Systems Europe Ltd. (Abingdon, Oxfordshire, UK); essentially fatty acid-free albumin from Sigma-Aldrich (St. Louis, MO, USA); α-modification of minimum essential medium (α-MEM) from Flow Laboratories (Irvine, Scotland, UK); [45 Ca]CaCl2 and Thermo Sequenase-TM II DYEnamic ET™ terminator cycle sequencing kit from Amersham Biosciences UK, Ltd., now part of GE Healthcare (Little Chalfont, Buckinghamshire, UK); HotStar Taq polymerase kit and QIAquick PCR purification kit from Qiagen Ltd. (Crawley, West Sussex, UK); culture dishes and multiwell plates from Costar, now part of Corning Life Sciences (Acton, MA, USA); radio-immunoassay (RIA) kit for prostaglandin E2 (PGE2 ) from Dupont-New England Nuclear Chemicals, now part of PerkinElmer Life and Analytical Sciences, Inc. (Waltham, MA, USA); enzyme-linked immunosorbent assay (ELISA) kits for RANKL and OPG from Biomedica Medizinprodukte GmbH & Co. KG (Vienna, Austria); TRIzol LS reagent, deoxyribonuclease I (amplification grade), and oligonucleotide primers from Invitrogen Ltd. (Paisley, Scotland, UK) or Applied Biosystems (Warrington, Cheshire, UK); kits for real-time polymerase chain reaction (PCR) analysis of FcRγ, DAP12, and OSCAR mRNA expression, fluorescence-labelled probes (reporter fluorescent dye VIC at the 5' end and quencher fluorescent dye TAMRA at the 3' end), and TaqMan Universal PCR Master Mix from Applied Biosystems; and the 1st Strand cDNA Synthesis Kit and PCR Core Kit from Roche Diagnostics GmbH (Mannheim, Germany). .. Synthetic salmon calcitonin was generously provided by Novartis International AG (Basel, Switzerland), 1,25(OH)2 -vitamin D3 (D3) by F. Hoffmann-La Roche Ltd. (Basel, Switzerland), and indomethacin by Merck Sharp & Dohme BV (Haarlem, The Netherlands).

    Article Title: Glucocorticoids employ the monomeric glucocorticoid receptor to potentiate vitamin D3 and parathyroid hormone–induced osteoclastogenesis
    Article Snippet: Materials Mouse OPG fused to human IgG1 Fc (OPG/Fc chimera), recombinant mouse M-CSF, recombinant extracellular domain of mouse RANKL (Arg72-Asp316; 462-TR), recombinant RANK/TNFRSF11A (RANKL neutralizing soluble RANK), and the ELISA kits for mouse RANKL and mouse OPG were purchased from R & D Systems (Minneapolis, MN, USA). .. HotStar Taq Polymerase Kit, QiaQuick PCR Purification Kit and RNeasy Mini Kit were from Qiagen (Hilden, Germany); DNA-Free and RNAqueous–4PCR Kit were obtained from Ambion (Austin, TX, USA); First-Strand cDNA Synthesis Kit and the PCR Core Kit were from Hoffmann-La Roche (Basel, Switzerland).

    Derivative Assay:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: The gene-specific primer (5′-ACCCAT-GAAGGTAAGAAGCCC-3′) used for PCR amplification of the target cDNA was based on the initially derived 5′ sequence of the N gene. .. PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing.

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing. .. Sequence data for the 2.5 kb product were obtained by initially using the diagnostic forward primer (GSP) and a poly-T sequencing primer, followed by primer-walking with newly designed 5 and 3 primers derived with the OLIGO 6 software (Molecular Biology Insights, Cascade, CO).

    Software:

    Article Title: Molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ECE) in ferrets.
    Article Snippet: PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 μl of cDNA template, 0.5 μM of the gene-specific primer, 2 μl of AUAP and the following cycling conditions: predenaturation at 95°C for 15 min, followed by 50 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, and extension at 72°C for 2 min and 30 s, and a final extension at 72°C for 7 min. PCR products of the expected size were purified for direct sequencing. .. Sequence data for the approximately 2 kb product were obtained by initially using the gene-specific primer described above and a poly-T sequencing primer, followed by primer-walking with newly designed 5′ and 3′ primers derived with the OLIGO 6 primer analysis software (Molecular Biology Insights, Cascade, CO).

    Article Title: Molecular characterization of noroviruses detected in diarrheic stools of Michigan and Wisconsin dairy calves: circulation of two distinct subgroups.
    Article Snippet: PCR was carried out with the QIAGEN HotStar Taq DNA Polymerase Kit, using 2 l of cDNA template, 0.5 M of the GSP, 2 l of AUAP and the following cycling conditions: pre-denaturation at 95 • C for 15 min, followed by 45 cycles of denaturation at 94 • C for 1 min, annealing at 55 • C for 1 min, and extension at 72 • C for 4 min, with a final extension of 72 • C for 10 min. PCR products of the expected size were extracted and purified from agarose gels with the QI-Aquick Gel Extraction Kit and submitted to GTSF-MSU for automated sequencing. .. Sequence data for the 2.5 kb product were obtained by initially using the diagnostic forward primer (GSP) and a poly-T sequencing primer, followed by primer-walking with newly designed 5 and 3 primers derived with the OLIGO 6 software (Molecular Biology Insights, Cascade, CO).

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