hotstar taq pcr buffer  (Qiagen)


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    Qiagen hotstar taq pcr buffer
    Hotstar Taq Pcr Buffer, supplied by Qiagen, used in various techniques. Bioz Stars score: 92/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hotstar taq pcr buffer/product/Qiagen
    Average 92 stars, based on 24 article reviews
    Price from $9.99 to $1999.99
    hotstar taq pcr buffer - by Bioz Stars, 2020-04
    92/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: Paragraph title: 2.7. Cloning and Sequencing ... The PCR reactions were carried out using 20 ng of bisulfite-treated DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol of forward and reverse PCR primers (Eurofins MWG Operon, Ebersberg, Germany) ( ) in a total volume of 25 μL.

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: .. The vector fragments containing the cloned PCR product were amplified using 3 μL DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol M13 forward and reverse primer provided within the kit in a total volume of 25 μL. .. The PCR program consisted of a denaturing step of 15 min at 95 °C followed by 35 cycles of 30 s at 95 °C, 30 s at 54 °C and 30 s at 72 °C, with a final extension of 7 min at 72 °C.

    Amplification:

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: A 267-bp fragment, −223 to +81 relative to the TSS, was amplified using primers not covering any CpGs ( and ). .. The PCR reactions were carried out using 20 ng of bisulfite-treated DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol of forward and reverse PCR primers (Eurofins MWG Operon, Ebersberg, Germany) ( ) in a total volume of 25 μL.

    Article Title: Simultaneous quantitative and allele-specific expression analysis with real competitive PCR
    Article Snippet: .. Step 1: PCR amplification Each PCR reaction contains 1 μL diluted cDNA (0.025 ng/μL), 0.5 μL 10× HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) and 2.14 μL ddH2 O. .. The PCR condition was: 95°C for 15 min for hot start, followed by denaturing at 94°C for 20 sec, annealing at 56°C for 30 sec and extension at 72°C for 1 min for 45 cycles, and finally incubated at 72°C for 3 min.

    Article Title: Linking Low-Level Stable Isotope Fractionation to Expression of the Cytochrome P450 Monooxygenase-Encoding ethB Gene for Elucidation of Methyl tert-Butyl Ether Biodegradation in Aerated Treatment Pond Systems ▿-Butyl Ether Biodegradation in Aerated Treatment Pond Systems ▿ †
    Article Snippet: The primer sets shown in Table were used to amplify genes encoding catabolic enzymes involved in MTBE degradation ( ethB , ethR , and mdpA ) from prepared cDNA samples and/or for sequencing of the obtained amplification products. .. A standard PCR mastermix contained 1× HotStar Taq PCR buffer, 1.5 mM MgCl2 , 0.2 mM each deoxynucleoside triphosphate, 0.5 μM each primer, 1.75 U HotStar Taq DNA polymerase (Qiagen GmbH, Germany), and 1 μl of template (cDNA or DNA).

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: .. The vector fragments containing the cloned PCR product were amplified using 3 μL DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol M13 forward and reverse primer provided within the kit in a total volume of 25 μL. .. The PCR program consisted of a denaturing step of 15 min at 95 °C followed by 35 cycles of 30 s at 95 °C, 30 s at 54 °C and 30 s at 72 °C, with a final extension of 7 min at 72 °C.

    Article Title: Using Progenitor Strain Information to Identify Quantitative Trait Nucleotides in Outbred Mice
    Article Snippet: Extension and amplification primers were designed using SpectroDesigner. .. A 5-μl PCR contained 2.5 ng of genomic DNA, 0.2 units of HotStar Taq , 5 pmol forward and reverse primers, 2 m m of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer [contains 1.5 m m MgCl2 , Tris-Cl, KCl, (NH4 )2 SO4 pH 8.7], and 25 m m MgCl2 (QIAGEN).

    Article Title: Novel IL10 gene family associations with systemic juvenile idiopathic arthritis
    Article Snippet: Primers designed by RealSNP assay (Sequenom) amplified approximately 100 bp of sequence surrounding the target SNP. .. PCR was carried out using 1× HotStar Taq PCR buffer 2.5 mM MgCl2 (Qiagen, Crawley, West Sussex, UK), 500 μM of each dNTP, 0.1 U Enzyme HotStar Taq polymerase (Qiagen), 100 nM primer, and 2.5 ng of genomic DNA in a total volume of 5 μl.

    Expressing:

    Article Title: Genome wide expression profiling of two accession of G. herbaceum L. in response to drought
    Article Snippet: Paragraph title: The Quantitative Gene Expression (QGE) analyses ... Each PCR reaction was performed with 1 μl diluted cDNA (0.025 ng/μl), 0.5 μL 10x HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) (primers list, Additional file ), and 2.14 μL ddH2O.

    Nucleic Acid Electrophoresis:

    Article Title: Linking Low-Level Stable Isotope Fractionation to Expression of the Cytochrome P450 Monooxygenase-Encoding ethB Gene for Elucidation of Methyl tert-Butyl Ether Biodegradation in Aerated Treatment Pond Systems ▿-Butyl Ether Biodegradation in Aerated Treatment Pond Systems ▿ †
    Article Snippet: A standard PCR mastermix contained 1× HotStar Taq PCR buffer, 1.5 mM MgCl2 , 0.2 mM each deoxynucleoside triphosphate, 0.5 μM each primer, 1.75 U HotStar Taq DNA polymerase (Qiagen GmbH, Germany), and 1 μl of template (cDNA or DNA). .. PCR product size was verified by gel electrophoresis on 1.2% agarose gels.

    Synthesized:

    Article Title: Commercially Available Outbred Mice for Genome-Wide Association Studies
    Article Snippet: .. Each 50 µl PCR contained 50 ng of genomic DNA, 1 Unit of HotStar Taq, 5 pmol of forward and reverse primers (synthesized at MWG Biotech, Ebersberg, Germany), 2 mM of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer (contains 1.5 mM MgCl2 , Tris-Cl, KCl and (NH4 )2 SO4 , pH 8.7) and 25 mM MgCl2 (Qiagen). .. We ran the PCR reactions using a Touchdown (TD) approach.

    Article Title: Using Progenitor Strain Information to Identify Quantitative Trait Nucleotides in Outbred Mice
    Article Snippet: Oligonucleotides were synthesized at Metabion (Martinsried, Germany). .. A 5-μl PCR contained 2.5 ng of genomic DNA, 0.2 units of HotStar Taq , 5 pmol forward and reverse primers, 2 m m of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer [contains 1.5 m m MgCl2 , Tris-Cl, KCl, (NH4 )2 SO4 pH 8.7], and 25 m m MgCl2 (QIAGEN).

    Size-exclusion Chromatography:

    Article Title: Simultaneous quantitative and allele-specific expression analysis with real competitive PCR
    Article Snippet: Step 1: PCR amplification Each PCR reaction contains 1 μL diluted cDNA (0.025 ng/μL), 0.5 μL 10× HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) and 2.14 μL ddH2 O. .. The PCR condition was: 95°C for 15 min for hot start, followed by denaturing at 94°C for 20 sec, annealing at 56°C for 30 sec and extension at 72°C for 1 min for 45 cycles, and finally incubated at 72°C for 3 min.

    Article Title: Genome wide expression profiling of two accession of G. herbaceum L. in response to drought
    Article Snippet: Each PCR reaction was performed with 1 μl diluted cDNA (0.025 ng/μl), 0.5 μL 10x HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) (primers list, Additional file ), and 2.14 μL ddH2O. .. The PCR condition was as follows: 95°C for 15 min for hot start, followed by denaturing at 94°C for 20 sec, annealing at 56°C for 30 sec, extension at 72°C for 1 min for 45 cycles, and finally, incubation at 72°C for 3 min.

    Article Title: Commercially Available Outbred Mice for Genome-Wide Association Studies
    Article Snippet: Each 50 µl PCR contained 50 ng of genomic DNA, 1 Unit of HotStar Taq, 5 pmol of forward and reverse primers (synthesized at MWG Biotech, Ebersberg, Germany), 2 mM of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer (contains 1.5 mM MgCl2 , Tris-Cl, KCl and (NH4 )2 SO4 , pH 8.7) and 25 mM MgCl2 (Qiagen). .. The temperature profile consisted of an initial enzyme activation at 95°C for 15 min, followed firstly by 13 cycles of 95°C for 30 sec, 64°C for 30 sec and 72°C for 60 sec, secondly by 29 cycles of 95°C for 30 sec, 57°C for 30 sec and 72°C for 60 sec, and finally by an incubation at 72°C for 7 min. PCR products were purified in a 96-well Millipore purification plate and resuspended in 30 µl of H2 O.

    Article Title: Using Progenitor Strain Information to Identify Quantitative Trait Nucleotides in Outbred Mice
    Article Snippet: A 5-μl PCR contained 2.5 ng of genomic DNA, 0.2 units of HotStar Taq , 5 pmol forward and reverse primers, 2 m m of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer [contains 1.5 m m MgCl2 , Tris-Cl, KCl, (NH4 )2 SO4 pH 8.7], and 25 m m MgCl2 (QIAGEN). .. The temperature profile consisted of an initial enzyme activation performed at 95° for 15 min, followed by 45 cycles of 94° for 20 sec, 56° for 30 sec, 72° for 60 sec, and a final incubation at 72° for 3 min. PCR products were treated with shrimp alkaline phosphatase (Sequenom) for 20 min at 37° to remove excess dNTPs.

    Purification:

    Article Title: Commercially Available Outbred Mice for Genome-Wide Association Studies
    Article Snippet: Each 50 µl PCR contained 50 ng of genomic DNA, 1 Unit of HotStar Taq, 5 pmol of forward and reverse primers (synthesized at MWG Biotech, Ebersberg, Germany), 2 mM of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer (contains 1.5 mM MgCl2 , Tris-Cl, KCl and (NH4 )2 SO4 , pH 8.7) and 25 mM MgCl2 (Qiagen). .. The temperature profile consisted of an initial enzyme activation at 95°C for 15 min, followed firstly by 13 cycles of 95°C for 30 sec, 64°C for 30 sec and 72°C for 60 sec, secondly by 29 cycles of 95°C for 30 sec, 57°C for 30 sec and 72°C for 60 sec, and finally by an incubation at 72°C for 7 min. PCR products were purified in a 96-well Millipore purification plate and resuspended in 30 µl of H2 O.

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: The vector fragments containing the cloned PCR product were amplified using 3 μL DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol M13 forward and reverse primer provided within the kit in a total volume of 25 μL. .. The sequencing reaction was carried out with 2 μL purified PCR product, 1× BigDye® Terminator v1.1 Sequencing buffer, 2 μL Big Dye® Terminator reaction mix v1.1 and 0.8 pmol M13 primer in a total volume of 10 μL.

    Article Title: Novel IL10 gene family associations with systemic juvenile idiopathic arthritis
    Article Snippet: PCR was carried out using 1× HotStar Taq PCR buffer 2.5 mM MgCl2 (Qiagen, Crawley, West Sussex, UK), 500 μM of each dNTP, 0.1 U Enzyme HotStar Taq polymerase (Qiagen), 100 nM primer, and 2.5 ng of genomic DNA in a total volume of 5 μl. .. Finally, the extended product was purified of excess ions using an ion-exchange resin.

    Activation Assay:

    Article Title: Commercially Available Outbred Mice for Genome-Wide Association Studies
    Article Snippet: Each 50 µl PCR contained 50 ng of genomic DNA, 1 Unit of HotStar Taq, 5 pmol of forward and reverse primers (synthesized at MWG Biotech, Ebersberg, Germany), 2 mM of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer (contains 1.5 mM MgCl2 , Tris-Cl, KCl and (NH4 )2 SO4 , pH 8.7) and 25 mM MgCl2 (Qiagen). .. The temperature profile consisted of an initial enzyme activation at 95°C for 15 min, followed firstly by 13 cycles of 95°C for 30 sec, 64°C for 30 sec and 72°C for 60 sec, secondly by 29 cycles of 95°C for 30 sec, 57°C for 30 sec and 72°C for 60 sec, and finally by an incubation at 72°C for 7 min. PCR products were purified in a 96-well Millipore purification plate and resuspended in 30 µl of H2 O.

    Article Title: Using Progenitor Strain Information to Identify Quantitative Trait Nucleotides in Outbred Mice
    Article Snippet: A 5-μl PCR contained 2.5 ng of genomic DNA, 0.2 units of HotStar Taq , 5 pmol forward and reverse primers, 2 m m of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer [contains 1.5 m m MgCl2 , Tris-Cl, KCl, (NH4 )2 SO4 pH 8.7], and 25 m m MgCl2 (QIAGEN). .. The temperature profile consisted of an initial enzyme activation performed at 95° for 15 min, followed by 45 cycles of 94° for 20 sec, 56° for 30 sec, 72° for 60 sec, and a final incubation at 72° for 3 min. PCR products were treated with shrimp alkaline phosphatase (Sequenom) for 20 min at 37° to remove excess dNTPs.

    Incubation:

    Article Title: Simultaneous quantitative and allele-specific expression analysis with real competitive PCR
    Article Snippet: Step 1: PCR amplification Each PCR reaction contains 1 μL diluted cDNA (0.025 ng/μL), 0.5 μL 10× HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) and 2.14 μL ddH2 O. .. The PCR condition was: 95°C for 15 min for hot start, followed by denaturing at 94°C for 20 sec, annealing at 56°C for 30 sec and extension at 72°C for 1 min for 45 cycles, and finally incubated at 72°C for 3 min.

    Article Title: Genome wide expression profiling of two accession of G. herbaceum L. in response to drought
    Article Snippet: Each PCR reaction was performed with 1 μl diluted cDNA (0.025 ng/μl), 0.5 μL 10x HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) (primers list, Additional file ), and 2.14 μL ddH2O. .. The PCR condition was as follows: 95°C for 15 min for hot start, followed by denaturing at 94°C for 20 sec, annealing at 56°C for 30 sec, extension at 72°C for 1 min for 45 cycles, and finally, incubation at 72°C for 3 min.

    Article Title: Commercially Available Outbred Mice for Genome-Wide Association Studies
    Article Snippet: Each 50 µl PCR contained 50 ng of genomic DNA, 1 Unit of HotStar Taq, 5 pmol of forward and reverse primers (synthesized at MWG Biotech, Ebersberg, Germany), 2 mM of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer (contains 1.5 mM MgCl2 , Tris-Cl, KCl and (NH4 )2 SO4 , pH 8.7) and 25 mM MgCl2 (Qiagen). .. The temperature profile consisted of an initial enzyme activation at 95°C for 15 min, followed firstly by 13 cycles of 95°C for 30 sec, 64°C for 30 sec and 72°C for 60 sec, secondly by 29 cycles of 95°C for 30 sec, 57°C for 30 sec and 72°C for 60 sec, and finally by an incubation at 72°C for 7 min. PCR products were purified in a 96-well Millipore purification plate and resuspended in 30 µl of H2 O.

    Article Title: Using Progenitor Strain Information to Identify Quantitative Trait Nucleotides in Outbred Mice
    Article Snippet: A 5-μl PCR contained 2.5 ng of genomic DNA, 0.2 units of HotStar Taq , 5 pmol forward and reverse primers, 2 m m of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer [contains 1.5 m m MgCl2 , Tris-Cl, KCl, (NH4 )2 SO4 pH 8.7], and 25 m m MgCl2 (QIAGEN). .. The temperature profile consisted of an initial enzyme activation performed at 95° for 15 min, followed by 45 cycles of 94° for 20 sec, 56° for 30 sec, 72° for 60 sec, and a final incubation at 72° for 3 min. PCR products were treated with shrimp alkaline phosphatase (Sequenom) for 20 min at 37° to remove excess dNTPs.

    Article Title: Novel IL10 gene family associations with systemic juvenile idiopathic arthritis
    Article Snippet: PCR was carried out using 1× HotStar Taq PCR buffer 2.5 mM MgCl2 (Qiagen, Crawley, West Sussex, UK), 500 μM of each dNTP, 0.1 U Enzyme HotStar Taq polymerase (Qiagen), 100 nM primer, and 2.5 ng of genomic DNA in a total volume of 5 μl. .. PCR was followed by incubation with shrimp alkaline phosphatase to digest unincorporated dNTPs and primers.

    Polymerase Chain Reaction:

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: .. The PCR reactions were carried out using 20 ng of bisulfite-treated DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol of forward and reverse PCR primers (Eurofins MWG Operon, Ebersberg, Germany) ( ) in a total volume of 25 μL. .. The PCR program consisted of a denaturing step of 15 min at 95 °C followed by 35 cycles of 30 s at 95 °C, 30 s at 54 °C and 30 s at 72 °C, with a final extension of 7 min at 72 °C.

    Article Title: Simultaneous quantitative and allele-specific expression analysis with real competitive PCR
    Article Snippet: .. Step 1: PCR amplification Each PCR reaction contains 1 μL diluted cDNA (0.025 ng/μL), 0.5 μL 10× HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) and 2.14 μL ddH2 O. .. The PCR condition was: 95°C for 15 min for hot start, followed by denaturing at 94°C for 20 sec, annealing at 56°C for 30 sec and extension at 72°C for 1 min for 45 cycles, and finally incubated at 72°C for 3 min.

    Article Title: Genome wide expression profiling of two accession of G. herbaceum L. in response to drought
    Article Snippet: .. Each PCR reaction was performed with 1 μl diluted cDNA (0.025 ng/μl), 0.5 μL 10x HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) (primers list, Additional file ), and 2.14 μL ddH2O. .. The PCR condition was as follows: 95°C for 15 min for hot start, followed by denaturing at 94°C for 20 sec, annealing at 56°C for 30 sec, extension at 72°C for 1 min for 45 cycles, and finally, incubation at 72°C for 3 min.

    Article Title: Commercially Available Outbred Mice for Genome-Wide Association Studies
    Article Snippet: .. Each 50 µl PCR contained 50 ng of genomic DNA, 1 Unit of HotStar Taq, 5 pmol of forward and reverse primers (synthesized at MWG Biotech, Ebersberg, Germany), 2 mM of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer (contains 1.5 mM MgCl2 , Tris-Cl, KCl and (NH4 )2 SO4 , pH 8.7) and 25 mM MgCl2 (Qiagen). .. We ran the PCR reactions using a Touchdown (TD) approach.

    Article Title: Linking Low-Level Stable Isotope Fractionation to Expression of the Cytochrome P450 Monooxygenase-Encoding ethB Gene for Elucidation of Methyl tert-Butyl Ether Biodegradation in Aerated Treatment Pond Systems ▿-Butyl Ether Biodegradation in Aerated Treatment Pond Systems ▿ †
    Article Snippet: .. A standard PCR mastermix contained 1× HotStar Taq PCR buffer, 1.5 mM MgCl2 , 0.2 mM each deoxynucleoside triphosphate, 0.5 μM each primer, 1.75 U HotStar Taq DNA polymerase (Qiagen GmbH, Germany), and 1 μl of template (cDNA or DNA). .. The conditions of the PCR were as follows: initial denaturation for 15 min at 95°C, 35 cycles of 94°C for 1 min, annealing (Table ) for 1 min, and 72°C for 1 min 30 s of extension, followed by a final elongation step of 20 min at 72°C.

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: .. The vector fragments containing the cloned PCR product were amplified using 3 μL DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol M13 forward and reverse primer provided within the kit in a total volume of 25 μL. .. The PCR program consisted of a denaturing step of 15 min at 95 °C followed by 35 cycles of 30 s at 95 °C, 30 s at 54 °C and 30 s at 72 °C, with a final extension of 7 min at 72 °C.

    Article Title: Using Progenitor Strain Information to Identify Quantitative Trait Nucleotides in Outbred Mice
    Article Snippet: .. A 5-μl PCR contained 2.5 ng of genomic DNA, 0.2 units of HotStar Taq , 5 pmol forward and reverse primers, 2 m m of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer [contains 1.5 m m MgCl2 , Tris-Cl, KCl, (NH4 )2 SO4 pH 8.7], and 25 m m MgCl2 (QIAGEN). .. The temperature profile consisted of an initial enzyme activation performed at 95° for 15 min, followed by 45 cycles of 94° for 20 sec, 56° for 30 sec, 72° for 60 sec, and a final incubation at 72° for 3 min. PCR products were treated with shrimp alkaline phosphatase (Sequenom) for 20 min at 37° to remove excess dNTPs.

    Article Title: Reticulate Evolution of the Rye Genome [W]Reticulate Evolution of the Rye Genome [W] [OPEN]
    Article Snippet: .. PCR was conducted in a total volume of 20 μL (20 ng of genomic DNA, 1× HotStar Taq PCR buffer, 250 nM each primer, 200 μM deoxynucleotide triphosphates, and 0.5 units of HotStar Taq DNA polymerase [Qiagen]). .. A touch-down PCR profile was applied (initial denaturation: 15 min at 95°C, 45 cycles: denaturation at 94°C for 1 min, annealing for 1 min [1°C incremental reduction from 65 to 55°C in the first 10 cycles and then 55°C] and extension at 72°C for 1 min [10 min at final extension]).

    Article Title: Novel IL10 gene family associations with systemic juvenile idiopathic arthritis
    Article Snippet: .. PCR was carried out using 1× HotStar Taq PCR buffer 2.5 mM MgCl2 (Qiagen, Crawley, West Sussex, UK), 500 μM of each dNTP, 0.1 U Enzyme HotStar Taq polymerase (Qiagen), 100 nM primer, and 2.5 ng of genomic DNA in a total volume of 5 μl. .. PCR was followed by incubation with shrimp alkaline phosphatase to digest unincorporated dNTPs and primers.

    Mass Spectrometry:

    Article Title: Genome wide expression profiling of two accession of G. herbaceum L. in response to drought
    Article Snippet: The Quantitative Gene Expression (QGE) analyses Recently, matrix-assisted lazer desorption ionization time of flight mass spectrometry (MALDI-TOF MS) was adopted for analyzing gene expression [ ]. .. Each PCR reaction was performed with 1 μl diluted cDNA (0.025 ng/μl), 0.5 μL 10x HotStar Taq PCR buffer, 0.2 μL MgCl2 (25 mM), 0.04 μL dNTP mix (25 mM each), 0.02 μL HotStar Taq Polymerase (50 U/μL, Qiagen), 0.1 μL competitor oligonucleotide (5 × 10-9 μM), 1 μL forward and reverse primer (1 μM each) (primers list, Additional file ), and 2.14 μL ddH2O.

    Sequencing:

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: Paragraph title: 2.7. Cloning and Sequencing ... The PCR reactions were carried out using 20 ng of bisulfite-treated DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol of forward and reverse PCR primers (Eurofins MWG Operon, Ebersberg, Germany) ( ) in a total volume of 25 μL.

    Article Title: Commercially Available Outbred Mice for Genome-Wide Association Studies
    Article Snippet: Paragraph title: Big dye sequencing ... Each 50 µl PCR contained 50 ng of genomic DNA, 1 Unit of HotStar Taq, 5 pmol of forward and reverse primers (synthesized at MWG Biotech, Ebersberg, Germany), 2 mM of each dNTP, 1× HotStar Taq PCR buffer as supplied by the enzyme manufacturer (contains 1.5 mM MgCl2 , Tris-Cl, KCl and (NH4 )2 SO4 , pH 8.7) and 25 mM MgCl2 (Qiagen).

    Article Title: Linking Low-Level Stable Isotope Fractionation to Expression of the Cytochrome P450 Monooxygenase-Encoding ethB Gene for Elucidation of Methyl tert-Butyl Ether Biodegradation in Aerated Treatment Pond Systems ▿-Butyl Ether Biodegradation in Aerated Treatment Pond Systems ▿ †
    Article Snippet: Paragraph title: PCR and sequencing reaction. ... A standard PCR mastermix contained 1× HotStar Taq PCR buffer, 1.5 mM MgCl2 , 0.2 mM each deoxynucleoside triphosphate, 0.5 μM each primer, 1.75 U HotStar Taq DNA polymerase (Qiagen GmbH, Germany), and 1 μl of template (cDNA or DNA).

    Article Title: Novel IL10 gene family associations with systemic juvenile idiopathic arthritis
    Article Snippet: Primers designed by RealSNP assay (Sequenom) amplified approximately 100 bp of sequence surrounding the target SNP. .. PCR was carried out using 1× HotStar Taq PCR buffer 2.5 mM MgCl2 (Qiagen, Crawley, West Sussex, UK), 500 μM of each dNTP, 0.1 U Enzyme HotStar Taq polymerase (Qiagen), 100 nM primer, and 2.5 ng of genomic DNA in a total volume of 5 μl.

    Transformation Assay:

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: The PCR reactions were carried out using 20 ng of bisulfite-treated DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol of forward and reverse PCR primers (Eurofins MWG Operon, Ebersberg, Germany) ( ) in a total volume of 25 μL. .. Three microliters of each PCR product were cloned into the pCR® 2.1-TOPO TA vector (Invitrogen), transformed into chemically-competent DH5α™-T1R Escherichia coli and grown overnight on LB plates containing 50 μg/mL ampicillin.

    Plasmid Preparation:

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: The PCR reactions were carried out using 20 ng of bisulfite-treated DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol of forward and reverse PCR primers (Eurofins MWG Operon, Ebersberg, Germany) ( ) in a total volume of 25 μL. .. Three microliters of each PCR product were cloned into the pCR® 2.1-TOPO TA vector (Invitrogen), transformed into chemically-competent DH5α™-T1R Escherichia coli and grown overnight on LB plates containing 50 μg/mL ampicillin.

    Article Title: Heterogeneous DNA Methylation Patterns in the GSTP1 Promoter Lead to Discordant Results between Assay Technologies and Impede Its Implementation as Epigenetic Biomarkers in Breast Cancer
    Article Snippet: .. The vector fragments containing the cloned PCR product were amplified using 3 μL DNA, 2 U HotStar Taq DNA Polymerase (Qiagen), 1× HotStar Taq PCR buffer supplemented with 1.6 mM MgCl2 (Qiagen), 125 μM dNTPs and 5 pmol M13 forward and reverse primer provided within the kit in a total volume of 25 μL. .. The PCR program consisted of a denaturing step of 15 min at 95 °C followed by 35 cycles of 30 s at 95 °C, 30 s at 54 °C and 30 s at 72 °C, with a final extension of 7 min at 72 °C.

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    Qiagen hotstart taq dna polymerase
    Hotstart Taq Dna Polymerase, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hotstart taq dna polymerase/product/Qiagen
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    Qiagen hotstar taq plus buffer
    Hotstar Taq Plus Buffer, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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