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    Structured Review

    Thermo Fisher dna polymerase
    Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 403 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna polymerase/product/Thermo Fisher
    Average 99 stars, based on 403 article reviews
    Price from $9.99 to $1999.99
    dna polymerase - by Bioz Stars, 2020-04
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    Related Articles

    Clone Assay:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Paragraph title: Generation of gene cluster DNA fragments and yeast assembly cloning in the yTREX vector ... Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Amplification:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Paragraph title: Sample procurement and RNA amplification: ... Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr.

    Article Title: Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rotaviruses: applications of the assay.
    Article Snippet: .. RT/PCR amplification The cDNA amplification reactions were carried out in 50 p.1 1 x PCR buffer (10 mM Tris-HCl pH 8.8) containing 1 pl of the RT reaction, 1.5mM MgCl , 50mM KCl, 0.1% Triton X-100, 2 mM of each c, 1 pM each primer and 2 units of "Dynazyme" DNA polymerase (Finnzymes Oy, Finland). .. Reaction mixtures were overlaid with 50 pl of light mineral oil (Sigma) and PCR was conducted in a thermal cycler (GenE, Techne) for 40 cycles (94°C for 1 min, 47°C for 2 min and 72°C for 2 miu) with a final 7-miu incubation at 72°C.

    Positive Control:

    Article Title: Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rotaviruses: applications of the assay.
    Article Snippet: RT/PCR amplification The cDNA amplification reactions were carried out in 50 p.1 1 x PCR buffer (10 mM Tris-HCl pH 8.8) containing 1 pl of the RT reaction, 1.5mM MgCl , 50mM KCl, 0.1% Triton X-100, 2 mM of each c, 1 pM each primer and 2 units of "Dynazyme" DNA polymerase (Finnzymes Oy, Finland). .. The positive control was viral RNA from SAl 1 rotavirus cultivated in MA-104 cells and extracted by the method of Herriug er al. (1982) .

    Construct:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Step 4: Verify correct assembly of the planned yTREX construct in silico using appropriate software (e.g. Clone Manager from Sci‐Ed Software), checking integrity of coding sequences, presence of RBSs and completeness of genes (start to stop codon). .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Electrophoresis:

    Article Title: Diagnostic performance of a rapid in-clinic test for the detection of Canine Parvovirus under different storage conditions and vaccination status.
    Article Snippet: Each 50 L reaction mixture contained PCR buffer 1× (KCl 50 mM, Tris-HCl 10 mM, pH 8.3), MgCl 2 2 mM, 200 mM of each deoxynucleotide, 1 M of each primer, 2 U of DNA Polymerase (Thermoscientific Maxima Hot Start Taq DNA polymerase) and 10 mL of template DNA. .. Following PCR, electrophoresis was performed using 8 L of the PCR products in a 2% Tris acetate-EDTA-agarose gel.

    Incubation:

    Article Title: Molecular Cloning and Functional Expression of the Potassium-Dependent Sodium–Calcium Exchanger from Human and Chicken Retinal Cone Photoreceptors
    Article Snippet: Incubation with Fab fragments from an anti-digoxigenin antibody (1:5000) from sheep, conjugated with alkaline phosphatase (Boehringer Mannheim, Indianapolis, IN) for 2 hr at 22°C was used for the detection of the digoxigenin-labeled riboprobes. .. Full-length chicken rod and cone NCKX and the alternative spliced cone NCKX cDNAs were cut out of their original vectors pBluescript with the restriction enzyme Eco RI, blunt-ended with DNA polymerase I, large (Klenow) fragment (Life Technologies), and gel-isolated (QIAquick Gel Extraction Kit, Qiagen).

    Article Title: HexaPrime: a novel method for detection of coronaviruses.
    Article Snippet: .. Five microliters of total RNA were mixed with 1.5 pM (1.5 l) of the reverse transcription (RT) primer (a list of RT primers used in the current study is given in Table 2 ), incubated at 65 • C for 5 min, cooled on ice for 2 min, and mixed with the RT MIX (25 U of Multi-Scribe Reverse Transcriptase (Life Technologies, Warsaw, Poland), 1 l of 10× DNA Polymerase I buffer (Thermo Scientific, Vilnius, Lithuania), 0.4 l of 100 mM dNTPs, 0.2 l of DMSO in a total volume of 3.5 l). .. Reactions were carried out for 120 min at 37 • C. Following the incubation, samples were heat-inactivated at 85 • C for 5 min.

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: .. Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr. .. The samples were blunt ended by addition of 2 μl of T4 DNA polymerase (Invitrogen) and incubation at 16° for 15 min. Ethanol precipitation was performed (70 μl 5 m NH4 OAc, 650 μl chilled 100% ethanol, 2 μl glycogen) and the sample was resuspended in 8 μl of RNase-free water.

    Article Title: Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rotaviruses: applications of the assay.
    Article Snippet: RT/PCR amplification The following RT/PCR method was used: 1 pl of dimethyl sulphoxide was added to 9 pl of the extracted RNA and heated at 97°C for 5 min to denature nucleic acids, cooled on ice, and the reaction buffer (6 pl 5 x AMV buffer [Promega]), 0.25 ul RNasin @omega), 3 p.l of 10 mM solutions of each of the deoxynucleoside triphosphates (dATP, dCTP, dGTP and dTTP), 2.5 pM of each primer, Beg9 and End9, and 10 units AMV reverse transcriptase (Promega) were added; reactions were incubated at room temperature for 10 min and subsequently at 42°C for 35 min. ending with 92°C for 2 min to inactivate the revem transcriptase. .. RT/PCR amplification The cDNA amplification reactions were carried out in 50 p.1 1 x PCR buffer (10 mM Tris-HCl pH 8.8) containing 1 pl of the RT reaction, 1.5mM MgCl , 50mM KCl, 0.1% Triton X-100, 2 mM of each c, 1 pM each primer and 2 units of "Dynazyme" DNA polymerase (Finnzymes Oy, Finland).

    Article Title: Diagnostic performance of a rapid in-clinic test for the detection of Canine Parvovirus under different storage conditions and vaccination status.
    Article Snippet: The aliquots were incubated at 65 • C for 10 min to inactivate PCR inhibitors and then they were chilled on ice (Uwatoko et al., 1995) . .. Each 50 L reaction mixture contained PCR buffer 1× (KCl 50 mM, Tris-HCl 10 mM, pH 8.3), MgCl 2 2 mM, 200 mM of each deoxynucleotide, 1 M of each primer, 2 U of DNA Polymerase (Thermoscientific Maxima Hot Start Taq DNA polymerase) and 10 mL of template DNA.

    Article Title: HexaPrime: a novel method for detection of coronaviruses.
    Article Snippet: Reactions were carried out for 120 min at 37 • C. Following the incubation, samples were heat-inactivated at 85 • C for 5 min. .. Subsequently, 5 l of the second-strand mix was added (0.5 U of RNase H, 0.5 l of 10× DNA Polymerase I buffer, 4.5 U of DNA Polymerase I (Thermo Scientific, Vilnius, Lithuania), 0.1 l of DMSO, and 3 pM of second-strand (SS) primer).

    In Silico:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Step 4: Verify correct assembly of the planned yTREX construct in silico using appropriate software (e.g. Clone Manager from Sci‐Ed Software), checking integrity of coding sequences, presence of RBSs and completeness of genes (start to stop codon). .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Expressing:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: The embryonic stem (ES) cell data sets were downloaded from the Gene Expression Omnibus (GEO) repository (unamplified samples 1–8; ) ( ). .. Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr.

    Flow Cytometry:

    Article Title: Tumor Necrosis Factor-Like Weak Inducer of Apoptosis-Induced Neurodegeneration
    Article Snippet: Loaded microbeads were placed into a flow cell forming a densely packed monolayer. .. Second-strand cDNA synthesis was performed by adding 40 U of DNA polymerase I, 2 U of RNase H, 10 U of Escherichia coli DNA ligase, and 0.5 m m each dNTP in 1× second-strand buffer (Invitrogen) in a final volume of 100 μl for 2 hr at 16°C.

    DNA Ligation:

    Article Title: Molecular Cloning and Functional Expression of the Potassium-Dependent Sodium–Calcium Exchanger from Human and Chicken Retinal Cone Photoreceptors
    Article Snippet: Full-length chicken rod and cone NCKX and the alternative spliced cone NCKX cDNAs were cut out of their original vectors pBluescript with the restriction enzyme Eco RI, blunt-ended with DNA polymerase I, large (Klenow) fragment (Life Technologies), and gel-isolated (QIAquick Gel Extraction Kit, Qiagen). .. Ligations were performed according to the manufacturer's protocol (Rapid DNA ligation kit, Boehringer Mannheim).

    Ligation:

    Article Title: Tumor Necrosis Factor-Like Weak Inducer of Apoptosis-Induced Neurodegeneration
    Article Snippet: Short sequences from the free template ends were obtained simultaneously by a fluorescence-based ligation-mediated sequencing method. .. Second-strand cDNA synthesis was performed by adding 40 U of DNA polymerase I, 2 U of RNase H, 10 U of Escherichia coli DNA ligase, and 0.5 m m each dNTP in 1× second-strand buffer (Invitrogen) in a final volume of 100 μl for 2 hr at 16°C.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rotaviruses: applications of the assay.
    Article Snippet: .. RT/PCR amplification The cDNA amplification reactions were carried out in 50 p.1 1 x PCR buffer (10 mM Tris-HCl pH 8.8) containing 1 pl of the RT reaction, 1.5mM MgCl , 50mM KCl, 0.1% Triton X-100, 2 mM of each c, 1 pM each primer and 2 units of "Dynazyme" DNA polymerase (Finnzymes Oy, Finland). .. Reaction mixtures were overlaid with 50 pl of light mineral oil (Sigma) and PCR was conducted in a thermal cycler (GenE, Techne) for 40 cycles (94°C for 1 min, 47°C for 2 min and 72°C for 2 miu) with a final 7-miu incubation at 72°C.

    Generated:

    Article Title: Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding.
    Article Snippet: Chinese CDC sequencing strategy The whole-genome sequences of 2019-nCoV from six samples (WH19001, WH19005, WH19002, WH19004, WH19008, and YS8011) were generated by a combination Sanger, Illumina, and Oxford nanopore sequencing. .. First, viral RNAs were extracted directly from clinical samples with the QIAamp Viral RNA Mini Kit, and then used to synthesise cDNA with the SuperScript III Reverse Transcriptase (ThermoFisher, Waltham, MA, USA) and N6 random primers, followed by second-strand synthesis with DNA Polymerase I, Large (Klenow) Fragment (ThermoFisher).

    Sequencing:

    Article Title: Molecular Cloning and Functional Expression of the Potassium-Dependent Sodium–Calcium Exchanger from Human and Chicken Retinal Cone Photoreceptors
    Article Snippet: Full-length chicken rod and cone NCKX and the alternative spliced cone NCKX cDNAs were cut out of their original vectors pBluescript with the restriction enzyme Eco RI, blunt-ended with DNA polymerase I, large (Klenow) fragment (Life Technologies), and gel-isolated (QIAquick Gel Extraction Kit, Qiagen). .. The orientation of the cDNAs was verified by sequencing vector/insert boundaries.

    Article Title: Tumor Necrosis Factor-Like Weak Inducer of Apoptosis-Induced Neurodegeneration
    Article Snippet: Signatures matching more than one gene (taking into account only nonexpressed sequence tags or expressed sequence tag European Molecular Biology Laboratory database entries) could be detected by clustering all matching sequences, excluding putative sequencing errors and sequence polymorphisms. .. Second-strand cDNA synthesis was performed by adding 40 U of DNA polymerase I, 2 U of RNase H, 10 U of Escherichia coli DNA ligase, and 0.5 m m each dNTP in 1× second-strand buffer (Invitrogen) in a final volume of 100 μl for 2 hr at 16°C.

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: An exemplary assembly scheme of a recombinant yTREX vector carrying the partial phz gene cluster from Pseudomonas aeruginosa and the β ‐galactosidase‐encoding lacZ reporter gene (for DNA sequence see Table ) including details on PCR primer sequences which contain the homologous regions for yeast recombineering is shown in Fig. . .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Article Title: Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding.
    Article Snippet: Paragraph title: Chinese CDC sequencing strategy ... First, viral RNAs were extracted directly from clinical samples with the QIAamp Viral RNA Mini Kit, and then used to synthesise cDNA with the SuperScript III Reverse Transcriptase (ThermoFisher, Waltham, MA, USA) and N6 random primers, followed by second-strand synthesis with DNA Polymerase I, Large (Klenow) Fragment (ThermoFisher).

    Recombinant:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: An exemplary assembly scheme of a recombinant yTREX vector carrying the partial phz gene cluster from Pseudomonas aeruginosa and the β ‐galactosidase‐encoding lacZ reporter gene (for DNA sequence see Table ) including details on PCR primer sequences which contain the homologous regions for yeast recombineering is shown in Fig. . .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Molecular Weight:

    Article Title: Diagnostic performance of a rapid in-clinic test for the detection of Canine Parvovirus under different storage conditions and vaccination status.
    Article Snippet: Each 50 L reaction mixture contained PCR buffer 1× (KCl 50 mM, Tris-HCl 10 mM, pH 8.3), MgCl 2 2 mM, 200 mM of each deoxynucleotide, 1 M of each primer, 2 U of DNA Polymerase (Thermoscientific Maxima Hot Start Taq DNA polymerase) and 10 mL of template DNA. .. Product sizes were determined using a 100 bp molecular weight ladder.

    Fluorescence:

    Article Title: Tumor Necrosis Factor-Like Weak Inducer of Apoptosis-Induced Neurodegeneration
    Article Snippet: Short sequences from the free template ends were obtained simultaneously by a fluorescence-based ligation-mediated sequencing method. .. Second-strand cDNA synthesis was performed by adding 40 U of DNA polymerase I, 2 U of RNase H, 10 U of Escherichia coli DNA ligase, and 0.5 m m each dNTP in 1× second-strand buffer (Invitrogen) in a final volume of 100 μl for 2 hr at 16°C.

    Isolation:

    Article Title: Tumor Necrosis Factor-Like Weak Inducer of Apoptosis-Induced Neurodegeneration
    Article Snippet: Second-strand cDNA synthesis was performed by adding 40 U of DNA polymerase I, 2 U of RNase H, 10 U of Escherichia coli DNA ligase, and 0.5 m m each dNTP in 1× second-strand buffer (Invitrogen) in a final volume of 100 μl for 2 hr at 16°C. .. Resuspended, double-stranded cDNA was digested with Dpn II; 3′ Dpn II fragments were isolated with streptavidin-coupled paramagnetic beads (Dynal Biotech, Hamburg, Germany) and released from the beads with a Bsm BI digest.

    Article Title: HexaPrime: a novel method for detection of coronaviruses.
    Article Snippet: HexaPrime reverse transcription and second-strand synthesis Total RNA that was isolated and purified as described in Section 2.3 was used for the synthesis of the first and second cDNA strands. .. Five microliters of total RNA were mixed with 1.5 pM (1.5 l) of the reverse transcription (RT) primer (a list of RT primers used in the current study is given in Table 2 ), incubated at 65 • C for 5 min, cooled on ice for 2 min, and mixed with the RT MIX (25 U of Multi-Scribe Reverse Transcriptase (Life Technologies, Warsaw, Poland), 1 l of 10× DNA Polymerase I buffer (Thermo Scientific, Vilnius, Lithuania), 0.4 l of 100 mM dNTPs, 0.2 l of DMSO in a total volume of 3.5 l).

    Labeling:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Adult mouse tissues were dissected from six week FVB mice and labeled per manufacturer's specifications without amplification. .. Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr.

    Mouse Assay:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: KitlSl /KitlSl-d male mice were purchased from the Jackson Laboratories. .. Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr.

    Polymerase Chain Reaction:

    Article Title: Assessment of genetic diversity in IL‐6 and RANTES promoters and their level in Saudi coronary artery disease patients
    Article Snippet: Paragraph title: 2.5. Polymerase chain reaction (PCR) ... Briefly, l μL of 10 pmol/L forward and reverse primers were added to the reaction mixture containing 12.5 μL of DNA polymerase (AmpliTaq Gold 360; Applied Biosystems, Foster city, CA, USA) and 2 μL of genomic DNA.

    Article Title: Molecular Cloning and Functional Expression of the Potassium-Dependent Sodium–Calcium Exchanger from Human and Chicken Retinal Cone Photoreceptors
    Article Snippet: Full-length chicken rod and cone NCKX and the alternative spliced cone NCKX cDNAs were cut out of their original vectors pBluescript with the restriction enzyme Eco RI, blunt-ended with DNA polymerase I, large (Klenow) fragment (Life Technologies), and gel-isolated (QIAquick Gel Extraction Kit, Qiagen). .. The vector pIE1/153A ( ) was digested with Sma I, treated with Calf Intestinal Alkaline Phosphatase (Life Technologies), and purified on PCR spin columns (Qiagen).

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH). .. Use a PCR programme according to the manufacturer's manual.

    Article Title: Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rotaviruses: applications of the assay.
    Article Snippet: .. RT/PCR amplification The cDNA amplification reactions were carried out in 50 p.1 1 x PCR buffer (10 mM Tris-HCl pH 8.8) containing 1 pl of the RT reaction, 1.5mM MgCl , 50mM KCl, 0.1% Triton X-100, 2 mM of each c, 1 pM each primer and 2 units of "Dynazyme" DNA polymerase (Finnzymes Oy, Finland). .. Reaction mixtures were overlaid with 50 pl of light mineral oil (Sigma) and PCR was conducted in a thermal cycler (GenE, Techne) for 40 cycles (94°C for 1 min, 47°C for 2 min and 72°C for 2 miu) with a final 7-miu incubation at 72°C.

    Article Title: Diagnostic performance of a rapid in-clinic test for the detection of Canine Parvovirus under different storage conditions and vaccination status.
    Article Snippet: .. Each 50 L reaction mixture contained PCR buffer 1× (KCl 50 mM, Tris-HCl 10 mM, pH 8.3), MgCl 2 2 mM, 200 mM of each deoxynucleotide, 1 M of each primer, 2 U of DNA Polymerase (Thermoscientific Maxima Hot Start Taq DNA polymerase) and 10 mL of template DNA. .. The thermal conditions of this protocol initially indicate an activation of Hot Start Taq DNA polymerase at 94 • C for10 min.

    Purification:

    Article Title: Molecular Cloning and Functional Expression of the Potassium-Dependent Sodium–Calcium Exchanger from Human and Chicken Retinal Cone Photoreceptors
    Article Snippet: Full-length chicken rod and cone NCKX and the alternative spliced cone NCKX cDNAs were cut out of their original vectors pBluescript with the restriction enzyme Eco RI, blunt-ended with DNA polymerase I, large (Klenow) fragment (Life Technologies), and gel-isolated (QIAquick Gel Extraction Kit, Qiagen). .. The vector pIE1/153A ( ) was digested with Sma I, treated with Calf Intestinal Alkaline Phosphatase (Life Technologies), and purified on PCR spin columns (Qiagen).

    Article Title: HexaPrime: a novel method for detection of coronaviruses.
    Article Snippet: HexaPrime reverse transcription and second-strand synthesis Total RNA that was isolated and purified as described in Section 2.3 was used for the synthesis of the first and second cDNA strands. .. Five microliters of total RNA were mixed with 1.5 pM (1.5 l) of the reverse transcription (RT) primer (a list of RT primers used in the current study is given in Table 2 ), incubated at 65 • C for 5 min, cooled on ice for 2 min, and mixed with the RT MIX (25 U of Multi-Scribe Reverse Transcriptase (Life Technologies, Warsaw, Poland), 1 l of 10× DNA Polymerase I buffer (Thermo Scientific, Vilnius, Lithuania), 0.4 l of 100 mM dNTPs, 0.2 l of DMSO in a total volume of 3.5 l).

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr. .. Samples were purified using RNeasy mini kit (QIAGEN), eluted in 50 μl H2 O, quantitated by OD260 , and 400 ng cRNA in 12 μl H2 O was subjected to a second round of amplification.

    Article Title: HexaPrime: a novel method for detection of coronaviruses.
    Article Snippet: HexaPrime reverse transcription and second-strand synthesis The resulting single-stranded cDNA was used for second-strand synthesis, with no purification step in-between. .. Subsequently, 5 l of the second-strand mix was added (0.5 U of RNase H, 0.5 l of 10× DNA Polymerase I buffer, 4.5 U of DNA Polymerase I (Thermo Scientific, Vilnius, Lithuania), 0.1 l of DMSO, and 3 pM of second-strand (SS) primer).

    Article Title: Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding.
    Article Snippet: First, viral RNAs were extracted directly from clinical samples with the QIAamp Viral RNA Mini Kit, and then used to synthesise cDNA with the SuperScript III Reverse Transcriptase (ThermoFisher, Waltham, MA, USA) and N6 random primers, followed by second-strand synthesis with DNA Polymerase I, Large (Klenow) Fragment (ThermoFisher). .. Viral cDNA libraries were prepared with use of the Nextera XT Library Prep Kit (Illumina, San Diego, CA, USA), then purified with Agencourt AMPure XP beads (Beckman Coulter, Brea, CA, USA), followed by quantification with an Invitrogen Qubit 2.0 Fluorometer.

    Plasmid Preparation:

    Article Title: Molecular Cloning and Functional Expression of the Potassium-Dependent Sodium–Calcium Exchanger from Human and Chicken Retinal Cone Photoreceptors
    Article Snippet: Full-length chicken rod and cone NCKX and the alternative spliced cone NCKX cDNAs were cut out of their original vectors pBluescript with the restriction enzyme Eco RI, blunt-ended with DNA polymerase I, large (Klenow) fragment (Life Technologies), and gel-isolated (QIAquick Gel Extraction Kit, Qiagen). .. The vector pIE1/153A ( ) was digested with Sma I, treated with Calf Intestinal Alkaline Phosphatase (Life Technologies), and purified on PCR spin columns (Qiagen).

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Paragraph title: Generation of gene cluster DNA fragments and yeast assembly cloning in the yTREX vector ... Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Software:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Step 4: Verify correct assembly of the planned yTREX construct in silico using appropriate software (e.g. Clone Manager from Sci‐Ed Software), checking integrity of coding sequences, presence of RBSs and completeness of genes (start to stop codon). .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Article Title: Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding.
    Article Snippet: First, viral RNAs were extracted directly from clinical samples with the QIAamp Viral RNA Mini Kit, and then used to synthesise cDNA with the SuperScript III Reverse Transcriptase (ThermoFisher, Waltham, MA, USA) and N6 random primers, followed by second-strand synthesis with DNA Polymerase I, Large (Klenow) Fragment (ThermoFisher). .. Chinese CDC sequencing strategy The raw fastQ files for each virus sample were filtered using previously described criteria, 18 then subjected to de novo assembly with the CLCBio software version 11.0.1.

    In Vitro:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr. .. In vitro transcription was performed with the MEGAscript high-yield transcription kit (Ambion) and incubated at 37° for 4 hr.

    Ethanol Precipitation:

    Article Title: Genomewide Discovery and Classification of Candidate Ovarian Fertility Genes in the Mouse
    Article Snippet: Second-strand synthesis was performed by adding 91 μl H2 O, 30 μl second-strand buffer, 3 μl 10 m m dNTPs, 1 μl Escherichia coli ligase, 4 μl DNA polymerase, 1 μl RNaseH (Invitrogen), and incubated at 16° for 2 hr. .. The samples were blunt ended by addition of 2 μl of T4 DNA polymerase (Invitrogen) and incubation at 16° for 15 min. Ethanol precipitation was performed (70 μl 5 m NH4 OAc, 650 μl chilled 100% ethanol, 2 μl glycogen) and the sample was resuspended in 8 μl of RNase-free water.

    Activation Assay:

    Article Title: Diagnostic performance of a rapid in-clinic test for the detection of Canine Parvovirus under different storage conditions and vaccination status.
    Article Snippet: Each 50 L reaction mixture contained PCR buffer 1× (KCl 50 mM, Tris-HCl 10 mM, pH 8.3), MgCl 2 2 mM, 200 mM of each deoxynucleotide, 1 M of each primer, 2 U of DNA Polymerase (Thermoscientific Maxima Hot Start Taq DNA polymerase) and 10 mL of template DNA. .. The thermal conditions of this protocol initially indicate an activation of Hot Start Taq DNA polymerase at 94 • C for10 min.

    DNA Purification:

    Article Title: Diagnostic performance of a rapid in-clinic test for the detection of Canine Parvovirus under different storage conditions and vaccination status.
    Article Snippet: A commercial DNA Purification kit (Thermoscientific Genomic DNA Purification Kit) was used to complete extraction from the specimens according to the manufacturer's protocol. .. Each 50 L reaction mixture contained PCR buffer 1× (KCl 50 mM, Tris-HCl 10 mM, pH 8.3), MgCl 2 2 mM, 200 mM of each deoxynucleotide, 1 M of each primer, 2 U of DNA Polymerase (Thermoscientific Maxima Hot Start Taq DNA polymerase) and 10 mL of template DNA.

    Gel Extraction:

    Article Title: Molecular Cloning and Functional Expression of the Potassium-Dependent Sodium–Calcium Exchanger from Human and Chicken Retinal Cone Photoreceptors
    Article Snippet: .. Full-length chicken rod and cone NCKX and the alternative spliced cone NCKX cDNAs were cut out of their original vectors pBluescript with the restriction enzyme Eco RI, blunt-ended with DNA polymerase I, large (Klenow) fragment (Life Technologies), and gel-isolated (QIAquick Gel Extraction Kit, Qiagen). .. The vector pIE1/153A ( ) was digested with Sma I, treated with Calf Intestinal Alkaline Phosphatase (Life Technologies), and purified on PCR spin columns (Qiagen).

    Homologous Recombination:

    Article Title: Protocols for yTREX/Tn5‐based gene cluster expression in Pseudomonas putida
    Article Snippet: Such homologies could result in false pairing during homologous recombination. .. Step 5: Amplify DNA fragments of the gene cluster of interest via PCR with homology arms using appropriate DNA comprising target genes as template, 0.1 μM of each primer and a DNA polymerase with a low error rate like Phusion High‐Fidelity DNA Polymerase (Thermo Fisher Scientific GmbH).

    Nanopore Sequencing:

    Article Title: Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding.
    Article Snippet: Chinese CDC sequencing strategy The whole-genome sequences of 2019-nCoV from six samples (WH19001, WH19005, WH19002, WH19004, WH19008, and YS8011) were generated by a combination Sanger, Illumina, and Oxford nanopore sequencing. .. First, viral RNAs were extracted directly from clinical samples with the QIAamp Viral RNA Mini Kit, and then used to synthesise cDNA with the SuperScript III Reverse Transcriptase (ThermoFisher, Waltham, MA, USA) and N6 random primers, followed by second-strand synthesis with DNA Polymerase I, Large (Klenow) Fragment (ThermoFisher).

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  • 99
    Thermo Fisher platinum superfi dna polymerase
    Clustering of samples amplified with two different polymerases on a non-metric multidimensional scaling (NMDS) plot of the Bray-Curtis dissimilarities. Points are colored by applied extraction kit. The encircled pairs correspond to a single sample where each data point represents one 16 S rRNA amplification with Phusion Hot Start II High-Fidelity <t>DNA</t> Polymerase and the another with Platinum <t>SuperFi</t> DNA Polymerase. Sample pairs labeled with * were stored in PBS.
    Platinum Superfi Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum superfi dna polymerase/product/Thermo Fisher
    Average 99 stars, based on 249 article reviews
    Price from $9.99 to $1999.99
    platinum superfi dna polymerase - by Bioz Stars, 2020-04
    99/100 stars
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    94
    Thermo Fisher dna polymerases
    Genotyping of Arabidopsis thaliana plants from SALK_100012 line with (a) <t>Phusion</t> High-Fidelity <t>DNA</t> polymerase; (b) Ex-Taq DNA polymerase; (c) Taq DNA polymerase; (d) Emerald Amp GT PCR Master Mix. WT - DNA from wild-type Arabidopsis plant (positive control for wild-type PCR band); 1–8 - individual plants from SALK_100012 line. Molecular weight DNA standards are shown on each side. Loading order for each DNA sample: PCR reaction for wild-type allele, PCR reaction for T-DNA mutant allele.
    Dna Polymerases, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna polymerases/product/Thermo Fisher
    Average 94 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    dna polymerases - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    Image Search Results


    Clustering of samples amplified with two different polymerases on a non-metric multidimensional scaling (NMDS) plot of the Bray-Curtis dissimilarities. Points are colored by applied extraction kit. The encircled pairs correspond to a single sample where each data point represents one 16 S rRNA amplification with Phusion Hot Start II High-Fidelity DNA Polymerase and the another with Platinum SuperFi DNA Polymerase. Sample pairs labeled with * were stored in PBS.

    Journal: Scientific Reports

    Article Title: The impact of storage buffer, DNA extraction method, and polymerase on microbial analysis

    doi: 10.1038/s41598-018-24573-y

    Figure Lengend Snippet: Clustering of samples amplified with two different polymerases on a non-metric multidimensional scaling (NMDS) plot of the Bray-Curtis dissimilarities. Points are colored by applied extraction kit. The encircled pairs correspond to a single sample where each data point represents one 16 S rRNA amplification with Phusion Hot Start II High-Fidelity DNA Polymerase and the another with Platinum SuperFi DNA Polymerase. Sample pairs labeled with * were stored in PBS.

    Article Snippet: Platinum SuperFi DNA Polymerase (Thermo Fisher Scientific) and the Phusion Hot Start II High-Fidelity DNA Polymerase (Thermo Fisher Scientific) were both tested for amplification.

    Techniques: Amplification, Labeling

    DNA sequencing of PTEN isolated from OVCAR-3 cells A. Electropherogram of the PTEN sequence highlighting the portion of cDNA bearing the mutation in OVCAR-3 (middle panel), as compared to the same region of cDNA from OAW42 cells, which express wild-type PTEN (left panel). The sequencing of the PTEN exon 5 of genomic DNA from OVCAR-3 (right panel) indicates that the mutation is effectively present in the germ line (i.e., it is not a PCR artefact) and that it does not occur in the paired allele. B. Localizationof the relevant functional domains in the molecule of PTEN. The mutated aminoacid position (155) in the phosphatase domain is indicated in red. C. Predicted structure models of WT and Y155C PTEN molecules in which the functional domains are marked in different colors: PIP2 Binding Domain (in green); Phosphatase domain (in blue); C2-domain (in orange); C-tail and PDZ domain (in magentas). In red, marked with an arrow, is indicated the tyrosine mutated in cysteine at position 155. D. Overlay of the wild-type (green) and Y155C mutated (red) phosphatase domain predicted structures. The Thyrosine to Cysteine substitution at position 155 (magentas) is pointed by the arrow. E. Predicted bonds between amino acids in the wild-type and in the Y155C mutated PTEN structure. a ) Tyr155 (red) in wild-type PTEN forms H bonds (green) with Gly127 and Lys128; b ) Cys155, in mutant PTEN forms a disulfide bridge with Cys136, which in wild-type PTEN would form an H-bond with Ala153.

    Journal: Oncotarget

    Article Title: PTEN dephosphorylates AKT to prevent the expression of GLUT1 on plasmamembrane and to limit glucose consumption in cancer cells

    doi: 10.18632/oncotarget.13113

    Figure Lengend Snippet: DNA sequencing of PTEN isolated from OVCAR-3 cells A. Electropherogram of the PTEN sequence highlighting the portion of cDNA bearing the mutation in OVCAR-3 (middle panel), as compared to the same region of cDNA from OAW42 cells, which express wild-type PTEN (left panel). The sequencing of the PTEN exon 5 of genomic DNA from OVCAR-3 (right panel) indicates that the mutation is effectively present in the germ line (i.e., it is not a PCR artefact) and that it does not occur in the paired allele. B. Localizationof the relevant functional domains in the molecule of PTEN. The mutated aminoacid position (155) in the phosphatase domain is indicated in red. C. Predicted structure models of WT and Y155C PTEN molecules in which the functional domains are marked in different colors: PIP2 Binding Domain (in green); Phosphatase domain (in blue); C2-domain (in orange); C-tail and PDZ domain (in magentas). In red, marked with an arrow, is indicated the tyrosine mutated in cysteine at position 155. D. Overlay of the wild-type (green) and Y155C mutated (red) phosphatase domain predicted structures. The Thyrosine to Cysteine substitution at position 155 (magentas) is pointed by the arrow. E. Predicted bonds between amino acids in the wild-type and in the Y155C mutated PTEN structure. a ) Tyr155 (red) in wild-type PTEN forms H bonds (green) with Gly127 and Lys128; b ) Cys155, in mutant PTEN forms a disulfide bridge with Cys136, which in wild-type PTEN would form an H-bond with Ala153.

    Article Snippet: PTEN cDNA was then cloned by PCR reaction employing the Platinum Pfx DNA polymerase (Thermo Scientific) with the following primers 5’-phosphorylated: forward 5’-CATTTCCATCCTGCAGAAGAAG-3’; reverse 5’-CCCAATACAGATTCACTTCCTTTAG-3’.

    Techniques: DNA Sequencing, Isolation, Sequencing, Mutagenesis, Polymerase Chain Reaction, Functional Assay, Binding Assay

    Genotyping of Arabidopsis thaliana plants from SALK_100012 line with (a) Phusion High-Fidelity DNA polymerase; (b) Ex-Taq DNA polymerase; (c) Taq DNA polymerase; (d) Emerald Amp GT PCR Master Mix. WT - DNA from wild-type Arabidopsis plant (positive control for wild-type PCR band); 1–8 - individual plants from SALK_100012 line. Molecular weight DNA standards are shown on each side. Loading order for each DNA sample: PCR reaction for wild-type allele, PCR reaction for T-DNA mutant allele.

    Journal: BioNanoScience

    Article Title: Selection of efficient Taq DNA polymerase to optimize T-DNA genotyping method for rapid detection of mutant Arabidopsis thaliana plants

    doi: 10.1007/s12668-016-0253-6

    Figure Lengend Snippet: Genotyping of Arabidopsis thaliana plants from SALK_100012 line with (a) Phusion High-Fidelity DNA polymerase; (b) Ex-Taq DNA polymerase; (c) Taq DNA polymerase; (d) Emerald Amp GT PCR Master Mix. WT - DNA from wild-type Arabidopsis plant (positive control for wild-type PCR band); 1–8 - individual plants from SALK_100012 line. Molecular weight DNA standards are shown on each side. Loading order for each DNA sample: PCR reaction for wild-type allele, PCR reaction for T-DNA mutant allele.

    Article Snippet: The following DNA polymerases were tested: Phusion High-Fidelity DNA polymerase (ThermoFisher Scientific, USA, cat #F530S), Ex-Taq DNA polymerase (Clontech, USA, cat #RR001A), Taq DNA polymerase (SibEnzyme, Russia, cat #E331), Emerald Amp GT PCR Master Mix (Clontech, USA, cat #RR310A).

    Techniques: Polymerase Chain Reaction, Positive Control, Molecular Weight, Mutagenesis

    Primase vs. primer-dependent DNA polymerase activity of intact PolpTN2, PolPTNΔ 311–923 and Taq polymerase. ( A ) Intact PolpTN2 (40 ng/µl), ( B ) PolpTN2Δ 311–923 (80 ng/µl) or ( C ) Taq DNA polymerase (0,05 u/µl) were incubated at 70°C in Taq buffer + 0,4 mM dNTP and 2 ng/µl M13mp18 DNA, which was or was not hybridized to the complementary oligonucleotide primer M13 forward ( Supplementary Table S1 ). At t = 0, 5, 10 and 15 min, aliquots were withdrawn and quenched into 25 mM EDTA. The amount of ds DNA synthesized was then determined using Sybr® Green I fluorescence as described in Materials and Methods. Circles, continuous line: synthesis with hybridized primer; squares, dashed line: synthesis without primer. Points are the average of two determinations. The standard deviation is indicated by error bars.

    Journal: Nucleic Acids Research

    Article Title: A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2

    doi: 10.1093/nar/gkt1385

    Figure Lengend Snippet: Primase vs. primer-dependent DNA polymerase activity of intact PolpTN2, PolPTNΔ 311–923 and Taq polymerase. ( A ) Intact PolpTN2 (40 ng/µl), ( B ) PolpTN2Δ 311–923 (80 ng/µl) or ( C ) Taq DNA polymerase (0,05 u/µl) were incubated at 70°C in Taq buffer + 0,4 mM dNTP and 2 ng/µl M13mp18 DNA, which was or was not hybridized to the complementary oligonucleotide primer M13 forward ( Supplementary Table S1 ). At t = 0, 5, 10 and 15 min, aliquots were withdrawn and quenched into 25 mM EDTA. The amount of ds DNA synthesized was then determined using Sybr® Green I fluorescence as described in Materials and Methods. Circles, continuous line: synthesis with hybridized primer; squares, dashed line: synthesis without primer. Points are the average of two determinations. The standard deviation is indicated by error bars.

    Article Snippet: Non-radioactive primase and DNA polymerase assays Reactions were carried out in Taq polymerase buffer (75 mM Tris-HCl pH 8.8 at 25°C, 20 mM (NH4 )2 SO4 , 0.01% Tween 20) (Thermo) containing 2.5 mM MgCl2 , 0.4 mM of each of the four dNTPs, 2 ng/µl of M13mp18 single-stranded DNA (New England Biolabs) and enzyme components as described in the text.

    Techniques: Activity Assay, Incubation, Synthesized, SYBR Green Assay, Fluorescence, Standard Deviation

    Efficiency of intact PolpTN2 and PolpTN2Δ 311–923 in priming DNA synthesis by T. nautilus PolB and Taq DNA polymerase. T. nautilus PolB (8 ng/µl) or Taq polymerase (0,05 u/µl) were incubated at 70°C in Taq buffer + 0,4 mM dNTP and 2 ng/µl M13mp18 DNA (without annealed primer) with increasing concentrations of either intact PolpTN2 ( A ) or PolpTN2 Δ 311–923 ( B ). DNA synthesis by intact PolpTN2 or PolpTN2 Δ 311–923 alone was included as a control. At t = 0 and 6 min, aliquots were withdrawn, quenched in 25 mM EDTA and assayed for ds DNA using Sybr® Green I fluorescence. Circles, continuous line: PolpTN2 or PolpTN2 Δ 311–923 alone; squares, dashed line: PolpTN2 or PolpTN2 Δ 311–923 plus Taq DNA polymerase; triangles, dotted line: PolpTN2 or PolpTN2 Δ 311–923 plus T. nautilus PolB DNA polymerase. Points are the average of three determinations. The standard deviation is indicated by error bars.

    Journal: Nucleic Acids Research

    Article Title: A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2

    doi: 10.1093/nar/gkt1385

    Figure Lengend Snippet: Efficiency of intact PolpTN2 and PolpTN2Δ 311–923 in priming DNA synthesis by T. nautilus PolB and Taq DNA polymerase. T. nautilus PolB (8 ng/µl) or Taq polymerase (0,05 u/µl) were incubated at 70°C in Taq buffer + 0,4 mM dNTP and 2 ng/µl M13mp18 DNA (without annealed primer) with increasing concentrations of either intact PolpTN2 ( A ) or PolpTN2 Δ 311–923 ( B ). DNA synthesis by intact PolpTN2 or PolpTN2 Δ 311–923 alone was included as a control. At t = 0 and 6 min, aliquots were withdrawn, quenched in 25 mM EDTA and assayed for ds DNA using Sybr® Green I fluorescence. Circles, continuous line: PolpTN2 or PolpTN2 Δ 311–923 alone; squares, dashed line: PolpTN2 or PolpTN2 Δ 311–923 plus Taq DNA polymerase; triangles, dotted line: PolpTN2 or PolpTN2 Δ 311–923 plus T. nautilus PolB DNA polymerase. Points are the average of three determinations. The standard deviation is indicated by error bars.

    Article Snippet: Non-radioactive primase and DNA polymerase assays Reactions were carried out in Taq polymerase buffer (75 mM Tris-HCl pH 8.8 at 25°C, 20 mM (NH4 )2 SO4 , 0.01% Tween 20) (Thermo) containing 2.5 mM MgCl2 , 0.4 mM of each of the four dNTPs, 2 ng/µl of M13mp18 single-stranded DNA (New England Biolabs) and enzyme components as described in the text.

    Techniques: DNA Synthesis, Incubation, SYBR Green Assay, Fluorescence, Standard Deviation