taq polymerase kit  (Thermo Fisher)


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    Name:
    Taq DNA Polymerase
    Description:
    Thermo Scientific Taq DNA Polymerase is a highly thermostable DNA polymerase from the thermophilic bacterium Thermus aquaticus The enzyme catalyzes 5 →3 synthesis of DNA has no detectable 3 →5 exonuclease proofreading activity and possesses low 5 →3 exonuclease activity In addition Taq DNA Polymerase exhibits deoxynucleotidyl transferase activity which frequently results in the addition of extra adenines at the 3 end of PCR products Recombinant Taq DNA Polymerase is the ideal tool for standard PCR of templates 5 kb or shorter Highlights• Thermostable half life is more than 40 min at 95°C• Generates PCR products with 3 dA overhangs• Supplied with two buffers 10X Taq Buffer with KCl and 10X Taq Buffer with NH4 2SO4 The latter allows for PCR at wide range of magnesium concentrations and decreases unspecific priming• Incorporates modified nucleotides e g biotin digoxigenin fluorescently labeled nucleotides Applications• Routine PCR amplification of DNA fragments up to 5 kb• High throughput PCR• DNA labelingNote• The error rate of Taq DNA Polymerase in PCR is 2 2 x 10 5 errors per nt per cycle Accordingly the accuracy of PCR is 4 5 x 104 Accuracy is an inverse of the error rate and shows an average number of correct nucleotides incorporated before an error occurs • The 10X Taq Buffer without Detergent is recommended for microarray experiments
    Catalog Number:
    ep0401
    Price:
    None
    Applications:
    PCR|PCR & Real-Time PCR|Routine PCR
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher taq polymerase kit
    Thermo Scientific Taq DNA Polymerase is a highly thermostable DNA polymerase from the thermophilic bacterium Thermus aquaticus The enzyme catalyzes 5 →3 synthesis of DNA has no detectable 3 →5 exonuclease proofreading activity and possesses low 5 →3 exonuclease activity In addition Taq DNA Polymerase exhibits deoxynucleotidyl transferase activity which frequently results in the addition of extra adenines at the 3 end of PCR products Recombinant Taq DNA Polymerase is the ideal tool for standard PCR of templates 5 kb or shorter Highlights• Thermostable half life is more than 40 min at 95°C• Generates PCR products with 3 dA overhangs• Supplied with two buffers 10X Taq Buffer with KCl and 10X Taq Buffer with NH4 2SO4 The latter allows for PCR at wide range of magnesium concentrations and decreases unspecific priming• Incorporates modified nucleotides e g biotin digoxigenin fluorescently labeled nucleotides Applications• Routine PCR amplification of DNA fragments up to 5 kb• High throughput PCR• DNA labelingNote• The error rate of Taq DNA Polymerase in PCR is 2 2 x 10 5 errors per nt per cycle Accordingly the accuracy of PCR is 4 5 x 104 Accuracy is an inverse of the error rate and shows an average number of correct nucleotides incorporated before an error occurs • The 10X Taq Buffer without Detergent is recommended for microarray experiments
    https://www.bioz.com/result/taq polymerase kit/product/Thermo Fisher
    Average 99 stars, based on 38 article reviews
    Price from $9.99 to $1999.99
    taq polymerase kit - by Bioz Stars, 2020-08
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase
    Article Snippet: .. Six histidine codons were introduced onto the N-terminus of the Taq DNA polymerase and Taq DNA pol/TBD gene and the recombinant genes were cloned in frame into the NdeI site of the pLEX vector (Invitrogen). .. 5′–3′ Exonuclease-deficient mutants which lack the first 235 amino acids of the N-terminal were likewise generated.

    Amplification:

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: .. Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA). .. The reaction conditions consisted of 30 cycles of 95 °C for 1 min, 55 °C for 1 min and 74 °C for 2 min. A semi-nested PCR using the amplified products at a 1:50 dilution, the internal primer ML561 and primer MU379 was performed using similar PCR conditions as described above.

    Article Title: A test of somatic mosaicism in the androgen receptor gene of Canada lynx (Lynx canadensis)
    Article Snippet: .. Amplification was conducted in a 10ul reaction containing deionized water (Invitrogen), 10X PCR Reaction Buffer (Invitrogen), 50 mM MgCl2 (Invitrogen), 100 mM dNTP solution (Invitrogen), 3 mg/mL BSA, 40uM forward and reverse primers (Integrated DNA Technologies) mentioned above (forward primers labeled with the fluorescent dye HEX), 0.0375U Invitrogen Taq DNA Polymerase, and 5 ng of DNA. .. The PCR reaction was run in a Bio-Rad DNA Engine Dyad and Dyad Disciple thermocycler under the following conditions: 94 °C for 15 min; followed by 29 cycles of 94 °C for 30 s, 52 °C for 1 min 30 s, and 72 °C for 1 min 30 s, and completed with a step of 60 °C for 45 min. Amplified samples were run on an 80 mL, 1.5 % agarose gel stained with ethidium bromide at 90 volts for 45 min, and visualized under ultraviolet light and to determine sex.

    Labeling:

    Article Title: A test of somatic mosaicism in the androgen receptor gene of Canada lynx (Lynx canadensis)
    Article Snippet: .. Amplification was conducted in a 10ul reaction containing deionized water (Invitrogen), 10X PCR Reaction Buffer (Invitrogen), 50 mM MgCl2 (Invitrogen), 100 mM dNTP solution (Invitrogen), 3 mg/mL BSA, 40uM forward and reverse primers (Integrated DNA Technologies) mentioned above (forward primers labeled with the fluorescent dye HEX), 0.0375U Invitrogen Taq DNA Polymerase, and 5 ng of DNA. .. The PCR reaction was run in a Bio-Rad DNA Engine Dyad and Dyad Disciple thermocycler under the following conditions: 94 °C for 15 min; followed by 29 cycles of 94 °C for 30 s, 52 °C for 1 min 30 s, and 72 °C for 1 min 30 s, and completed with a step of 60 °C for 45 min. Amplified samples were run on an 80 mL, 1.5 % agarose gel stained with ethidium bromide at 90 volts for 45 min, and visualized under ultraviolet light and to determine sex.

    other:

    Article Title: Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans—Expression and characterization
    Article Snippet: The processivity values determined with the use of a heparin trap were much lower than the values previously published for the Taq DNA polymerase [ , ] and were within a range of 80 nt to 160 nt.

    Polymerase Chain Reaction:

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: .. Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA). .. The reaction conditions consisted of 30 cycles of 95 °C for 1 min, 55 °C for 1 min and 74 °C for 2 min. A semi-nested PCR using the amplified products at a 1:50 dilution, the internal primer ML561 and primer MU379 was performed using similar PCR conditions as described above.

    Article Title: Conditional mutagenesis by oligonucleotide-mediated integration of loxP sites in zebrafish
    Article Snippet: .. All PCR reactions were performed using either NEB 2X Taq Master Mix (M0270L), Thermo Scientific Taq (EP0402 and 2x PCR Master Mix Cat# AB-0575/DC), or Kapa 2G Fast ReadyMix +dye (Kapa Biosystems-KM5101). .. CRISPR activity was assessed by either direct sequencing of PCR amplicons, T7 assay (NEB M0302S), Surveyor assay (IDT 706025), or RFLP.

    Article Title: A test of somatic mosaicism in the androgen receptor gene of Canada lynx (Lynx canadensis)
    Article Snippet: .. Amplification was conducted in a 10ul reaction containing deionized water (Invitrogen), 10X PCR Reaction Buffer (Invitrogen), 50 mM MgCl2 (Invitrogen), 100 mM dNTP solution (Invitrogen), 3 mg/mL BSA, 40uM forward and reverse primers (Integrated DNA Technologies) mentioned above (forward primers labeled with the fluorescent dye HEX), 0.0375U Invitrogen Taq DNA Polymerase, and 5 ng of DNA. .. The PCR reaction was run in a Bio-Rad DNA Engine Dyad and Dyad Disciple thermocycler under the following conditions: 94 °C for 15 min; followed by 29 cycles of 94 °C for 30 s, 52 °C for 1 min 30 s, and 72 °C for 1 min 30 s, and completed with a step of 60 °C for 45 min. Amplified samples were run on an 80 mL, 1.5 % agarose gel stained with ethidium bromide at 90 volts for 45 min, and visualized under ultraviolet light and to determine sex.

    Article Title: Activation of alternative Jdp2 promoters and functional protein isoforms in T-cell lymphomas by retroviral insertion mutagenesis
    Article Snippet: .. To look for alternative splicing between published exons 1a through 1d, PCR reactions were done with forward primers Exon1a-154 (5′-tgggcaccgcgcctgcagcag-3′), Exon1b-148 (5′-ggaggagcgcgagcat-3′), Exon1c-70 (5′-gctctggctgggttaggagggaac-3′) or Exon1d-150 (5′-cagctgcctctctccatctt-3′) and reverse primer Intron2-87 (5′-tccttcgctcttcttcctcgtctagctt-3′) using 1/500 of the cDNA (corresponding to 4 ng of total RNA) per PCR reaction (Taq polymerase, Invitrogen). .. Each PCR reaction consisted of an initial melting step at 95°C for 5 min followed by 35 cycles of amplification, each consisting of a 30 s melting step at 95°C, annealing at 55–60°C for 30 s and elongation at 72°C for 45 s. A final elongation step at 72°C was done for 10 min. Quantitative real-time PCR (QRT-PCR) was done using Platinum SYBR Green QRT-PCR SuperMix UDG (Invitrogen) following the manufacturer's recommendations.

    Recombinant:

    Article Title: Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase
    Article Snippet: .. Six histidine codons were introduced onto the N-terminus of the Taq DNA polymerase and Taq DNA pol/TBD gene and the recombinant genes were cloned in frame into the NdeI site of the pLEX vector (Invitrogen). .. 5′–3′ Exonuclease-deficient mutants which lack the first 235 amino acids of the N-terminal were likewise generated.

    Plasmid Preparation:

    Article Title: Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase
    Article Snippet: .. Six histidine codons were introduced onto the N-terminus of the Taq DNA polymerase and Taq DNA pol/TBD gene and the recombinant genes were cloned in frame into the NdeI site of the pLEX vector (Invitrogen). .. 5′–3′ Exonuclease-deficient mutants which lack the first 235 amino acids of the N-terminal were likewise generated.

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    Thermo Fisher platinum taq dna polymerase high fidelity
    Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by <t>DNA</t> sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the <t>Taq</t> DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.
    Platinum Taq Dna Polymerase High Fidelity, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 894 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum taq dna polymerase high fidelity/product/Thermo Fisher
    Average 99 stars, based on 894 article reviews
    Price from $9.99 to $1999.99
    platinum taq dna polymerase high fidelity - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher superscript iii rt pcr platinum taq high fidelity kit
    ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all <t>three</t> of the cell populations were subjected to <t>RT-PCR</t> to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).
    Superscript Iii Rt Pcr Platinum Taq High Fidelity Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii rt pcr platinum taq high fidelity kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superscript iii rt pcr platinum taq high fidelity kit - by Bioz Stars, 2020-08
    99/100 stars
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    Image Search Results


    Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by DNA sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the Taq DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.

    Journal: BioMed Research International

    Article Title: LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity

    doi: 10.1155/2018/5657085

    Figure Lengend Snippet: Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by DNA sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the Taq DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.

    Article Snippet: PCR fragments of LraI operon amplified using Platinum™ Taq DNA Polymerase High Fidelity were cloned into pAZIL vector predigested with Sma I.

    Techniques: DNA Sequencing, Sequencing, Activity Assay

    Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by DNA sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the Taq DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.

    Journal: BioMed Research International

    Article Title: LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity

    doi: 10.1155/2018/5657085

    Figure Lengend Snippet: Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by DNA sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the Taq DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.

    Article Snippet: Platinum™ Taq DNA Polymerase High Fidelity (Thermo Fisher Scientific, Waltham, MA, USA) was used to amplify DNA fragments by PCR in GeneAmp PCR system 2700 thermal cycler (Applied Biosystems, Foster City, CA, USA).

    Techniques: DNA Sequencing, Sequencing, Activity Assay

    ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all three of the cell populations were subjected to RT-PCR to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).

    Journal: Journal of Virology

    Article Title: Transposon Mutagenesis of the Zika Virus Genome Highlights Regions Essential for RNA Replication and Restricted for Immune Evasion

    doi: 10.1128/JVI.00698-17

    Figure Lengend Snippet: ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all three of the cell populations were subjected to RT-PCR to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).

    Article Snippet: The viral RNA was then amplified in three segments by utilizing a SuperScript III RT-PCR Platinum Taq High Fidelity kit (Thermo Fisher Scientific).

    Techniques: Mutagenesis, Library Screening, Sequencing, Transfection, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Infection