longamp taq dna polymerase  (New England Biolabs)


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    Structured Review

    New England Biolabs longamp taq dna polymerase
    Longamp Taq Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/longamp taq dna polymerase/product/New England Biolabs
    Average 99 stars, based on 120 article reviews
    Price from $9.99 to $1999.99
    longamp taq dna polymerase - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Molecular Cloning:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Paragraph title: Sequence identification and molecular cloning ... Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc).

    Amplification:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: .. Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc). .. 5 0 and 3 0 RACE were performed using a GeneRACER kit (Invitrogen).

    Clone Assay:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc). .. PCR and RACE products were cloned into the pCR4-TOPO vector using the TOPO TA Cloning Kit for sequencing (Invitrogen).

    TA Cloning:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc). .. PCR and RACE products were cloned into the pCR4-TOPO vector using the TOPO TA Cloning Kit for sequencing (Invitrogen).

    Sequencing:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Paragraph title: Sequence identification and molecular cloning ... Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc).

    DNA Sequencing:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc). .. M13 forward and M13 reverse universal sequencing primers were employed for DNA sequencing using a BigDye Terminator Cycle Sequencing Kit v3.1 (Applied Biosystems) and an Applied Biosystems 3130 XL Genetic Analyser.

    Polymerase Chain Reaction:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Primers for Rapid Amplification of cDNA ends (RACE) and PCR were designed using Clone Manager 9.0 (Sci-Ed Software). .. Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc).

    Rapid Amplification of cDNA Ends:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Primers for Rapid Amplification of cDNA ends (RACE) and PCR were designed using Clone Manager 9.0 (Sci-Ed Software). .. Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc).

    Plasmid Preparation:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc). .. PCR and RACE products were cloned into the pCR4-TOPO vector using the TOPO TA Cloning Kit for sequencing (Invitrogen).

    Software:

    Article Title: Molecular characterisation of RIG-I-like helicases in the black flying fox, Pteropus alecto.
    Article Snippet: Primers for Rapid Amplification of cDNA ends (RACE) and PCR were designed using Clone Manager 9.0 (Sci-Ed Software). .. Full-length amplification was performed using LongAmp Taq DNA polymerase (New England BioLabs Inc).

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    New England Biolabs longamp taq dna polymerase
    Thermal shock is crucial for successful PCR amplification from fungal spores. A. fumigatus spores at three different concentrations (8 × 10 8 /ml, 1 × 10 8 /ml, and 5 × 10 7 /ml) were prepared as described in Basic Protocol 1 . 3 µl of spore suspension were used as the <t>DNA</t> template for PCR. ( A ) PCR result from spore suspension subjected to thermal shock of 95°C for 15 min and −80°C for 10 min prior to PCR, as described in Basic Protocol 2 . No thermal shock was done for spore suspensions from Panels B and C . Subsequently, a PCR was run with an initial denaturation step of 95°C for 1 min (A), 5 min (B), or 15 min (C) with primers ITS1 and D2 (expected PCR product ∼1.2 kb) and <t>LongAmp</t> <t>Taq</t> DNA polymerase with PCR conditions described in Basic Protocol 2 . P: positive PCR control amplified from genomic DNA (50 ng) of the A. fumigatus wild‐type strain; N: negative control (no DNA).
    Longamp Taq Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/longamp taq dna polymerase/product/New England Biolabs
    Average 99 stars, based on 123 article reviews
    Price from $9.99 to $1999.99
    longamp taq dna polymerase - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    94
    New England Biolabs neb longamp taq
    ssDNA production by aPCR. ( a ) aPCR reactions were assembled with a 50-molar excess of a forward primer for the amplification of a 1,000 nt ssDNA fragment using the M13mp18 ssDNA plasmid as template, and with 10 different polymerases that were tested for highest yield of ssDNA production (upper band: expected dsDNA size is 1,000 bp; lower band: expected ssDNA size is 1,000 nt) as judged by agarose gel electrophoresis (right panel). QuantaBio AccuStart HiFi, polymerase (lane 2, boxed) produced the highest amount without overlapping dsDNA contaminants. 1. Accustart; 2. Accustart HiFi; 3. Accustart II; 4. AccuPrime; 5. GoTaq; 6. DreamTaq; 7. Phusion; 8. Platinum SuperFi; 9. Q5; 10. Tth polymerase. ( b ) Biochemical validation of ssDNA production by incubating 1,000 nt aPCR reaction products with the ssDNA-specific ExoI or S1 nucleases or dsDNA-specific restriction enzymes Eco RI and Nae I (left panel). Agarose gel electrophoresis of the digestion products as labeled by lane (right panel). M: Marker, C: aPCR product control, ExoI: exonuclease I, S1: S1 nuclease, Enz: Eco RI + Nae I. ( c ) <t>NEB</t> <t>LongAmp</t> was used to generate ssDNA up to 15,000 nt long using lambda phage dsDNA as template. Purification of the 10 kb fragment shows a single band of higher molecular weight than the M13mp18 ssDNA (7,249 nt). ( d ) The primer design algorithm aPrime was used to select primers for product sizes between 500 and 3,000 nt using M13mp18 ssDNA as template and the Quantabio Accustart HiFi enzyme. SYBR Safe stained agarose gels illuminated under blue light show dsDNA as yellow bands, while ssDNA show as orange bands.
    Neb Longamp Taq, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/neb longamp taq/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    neb longamp taq - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

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    Thermal shock is crucial for successful PCR amplification from fungal spores. A. fumigatus spores at three different concentrations (8 × 10 8 /ml, 1 × 10 8 /ml, and 5 × 10 7 /ml) were prepared as described in Basic Protocol 1 . 3 µl of spore suspension were used as the DNA template for PCR. ( A ) PCR result from spore suspension subjected to thermal shock of 95°C for 15 min and −80°C for 10 min prior to PCR, as described in Basic Protocol 2 . No thermal shock was done for spore suspensions from Panels B and C . Subsequently, a PCR was run with an initial denaturation step of 95°C for 1 min (A), 5 min (B), or 15 min (C) with primers ITS1 and D2 (expected PCR product ∼1.2 kb) and LongAmp Taq DNA polymerase with PCR conditions described in Basic Protocol 2 . P: positive PCR control amplified from genomic DNA (50 ng) of the A. fumigatus wild‐type strain; N: negative control (no DNA).

    Journal: Current Protocols in Microbiology

    Article Title: Fast and Reliable PCR Amplification from Aspergillus fumigatus Spore Suspension Without Traditional DNA Extraction). Fast and reliable PCR amplification from Aspergillus fumigatus spore suspension without traditional DNA extraction

    doi: 10.1002/cpmc.89

    Figure Lengend Snippet: Thermal shock is crucial for successful PCR amplification from fungal spores. A. fumigatus spores at three different concentrations (8 × 10 8 /ml, 1 × 10 8 /ml, and 5 × 10 7 /ml) were prepared as described in Basic Protocol 1 . 3 µl of spore suspension were used as the DNA template for PCR. ( A ) PCR result from spore suspension subjected to thermal shock of 95°C for 15 min and −80°C for 10 min prior to PCR, as described in Basic Protocol 2 . No thermal shock was done for spore suspensions from Panels B and C . Subsequently, a PCR was run with an initial denaturation step of 95°C for 1 min (A), 5 min (B), or 15 min (C) with primers ITS1 and D2 (expected PCR product ∼1.2 kb) and LongAmp Taq DNA polymerase with PCR conditions described in Basic Protocol 2 . P: positive PCR control amplified from genomic DNA (50 ng) of the A. fumigatus wild‐type strain; N: negative control (no DNA).

    Article Snippet: Materials Spore suspension (Basic Protocol ) Primers (5 µM each, forward and reverse; experiment specific) LongAmp Taq DNA polymerase with 5× reaction buffer (New England Biolabs; M0323) dNTP mix (5 mM of each dNTP) Sterile, molecular‐grade water For agarose gel electrophoresis (also see Current Protocols article: Voytas, ): 6× DNA loading dye 1× TAE buffer (see recipe) Ethidium bromide or other DNA gel stain 0.2‐ml PCR tubes, 0.2‐ml/well 96‐well PCR plates, or 150 µl/well 384 well PCR plates with tight (preferably aluminum) seals Thermal cycler Centrifuge UV transilluminator Additional reagents and equipment for agarose gel electrophoresis (see Current Protocols article: Voytas, )

    Techniques: Polymerase Chain Reaction, Amplification, Negative Control

    Spore PCR using the supernatant from spore suspensions of different filamentous fungi with primers ITS1/D2 (expected PCR band size is ∼1.2 kb) and ITS1/ITS4 (expected PCR band size ∼600 bp). Two different spore concentrations were tested (i.e., 5 × 10 7 /ml and 1 × 10 7 /ml). 1 µl of the supernatant was used in the PCR reaction with the LongAmp Taq DNA polymerase. Positive PCR controls were amplified from DNA (50 ng) of the A. fumigatus wild‐type strain with primers ITS1/D2 (P1) and ITS1/ITS4 (P2). N: negative control (no DNA).

    Journal: Current Protocols in Microbiology

    Article Title: Fast and Reliable PCR Amplification from Aspergillus fumigatus Spore Suspension Without Traditional DNA Extraction). Fast and reliable PCR amplification from Aspergillus fumigatus spore suspension without traditional DNA extraction

    doi: 10.1002/cpmc.89

    Figure Lengend Snippet: Spore PCR using the supernatant from spore suspensions of different filamentous fungi with primers ITS1/D2 (expected PCR band size is ∼1.2 kb) and ITS1/ITS4 (expected PCR band size ∼600 bp). Two different spore concentrations were tested (i.e., 5 × 10 7 /ml and 1 × 10 7 /ml). 1 µl of the supernatant was used in the PCR reaction with the LongAmp Taq DNA polymerase. Positive PCR controls were amplified from DNA (50 ng) of the A. fumigatus wild‐type strain with primers ITS1/D2 (P1) and ITS1/ITS4 (P2). N: negative control (no DNA).

    Article Snippet: Materials Spore suspension (Basic Protocol ) Primers (5 µM each, forward and reverse; experiment specific) LongAmp Taq DNA polymerase with 5× reaction buffer (New England Biolabs; M0323) dNTP mix (5 mM of each dNTP) Sterile, molecular‐grade water For agarose gel electrophoresis (also see Current Protocols article: Voytas, ): 6× DNA loading dye 1× TAE buffer (see recipe) Ethidium bromide or other DNA gel stain 0.2‐ml PCR tubes, 0.2‐ml/well 96‐well PCR plates, or 150 µl/well 384 well PCR plates with tight (preferably aluminum) seals Thermal cycler Centrifuge UV transilluminator Additional reagents and equipment for agarose gel electrophoresis (see Current Protocols article: Voytas, )

    Techniques: Polymerase Chain Reaction, Amplification, Negative Control

    Derivative dissociation plots produced by different reagents using one microliter of lysate produced by homogenizing one H. armigera leg with 24 H. zea legs in 625 μl of modified squish buffer. (A) Amplification reactions made with KAPA SYBR Fast Master Mix (purple) and NEB LongAmp Taq polymerase and buffer with 2.5 mM MgCl2 (Red). Peaks representing 18S rRNA control amplicon and species-specific amplicons of H. armigera and H. zea are present in all reactions, but melting temperatures were different in two reagents. (B) An enlarged view of the derivative dissociation plot produced by amplification reactions with NEB LongAmp Taq polymerase and buffer. Although variations in dissociation plots between samples were observed, the 3-peak dissociation plot was clearly identifiable.

    Journal: Journal of Insect Science

    Article Title: Rapid Identification of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) Using Ribosomal RNA Internal Transcribed Spacer 1

    doi: 10.1093/jisesa/iev137

    Figure Lengend Snippet: Derivative dissociation plots produced by different reagents using one microliter of lysate produced by homogenizing one H. armigera leg with 24 H. zea legs in 625 μl of modified squish buffer. (A) Amplification reactions made with KAPA SYBR Fast Master Mix (purple) and NEB LongAmp Taq polymerase and buffer with 2.5 mM MgCl2 (Red). Peaks representing 18S rRNA control amplicon and species-specific amplicons of H. armigera and H. zea are present in all reactions, but melting temperatures were different in two reagents. (B) An enlarged view of the derivative dissociation plot produced by amplification reactions with NEB LongAmp Taq polymerase and buffer. Although variations in dissociation plots between samples were observed, the 3-peak dissociation plot was clearly identifiable.

    Article Snippet: Initial tests of species-specific fragment amplification were carried out using Crimson LongAmp Taq polymerase with a 45 s initial denaturation step at 95°C followed by 35 cycles of 15 s denaturation at 95°C, 10 s annealing at 60°C, and 30 s extension at 72°C.

    Techniques: Produced, Modification, Amplification

    ssDNA production by aPCR. ( a ) aPCR reactions were assembled with a 50-molar excess of a forward primer for the amplification of a 1,000 nt ssDNA fragment using the M13mp18 ssDNA plasmid as template, and with 10 different polymerases that were tested for highest yield of ssDNA production (upper band: expected dsDNA size is 1,000 bp; lower band: expected ssDNA size is 1,000 nt) as judged by agarose gel electrophoresis (right panel). QuantaBio AccuStart HiFi, polymerase (lane 2, boxed) produced the highest amount without overlapping dsDNA contaminants. 1. Accustart; 2. Accustart HiFi; 3. Accustart II; 4. AccuPrime; 5. GoTaq; 6. DreamTaq; 7. Phusion; 8. Platinum SuperFi; 9. Q5; 10. Tth polymerase. ( b ) Biochemical validation of ssDNA production by incubating 1,000 nt aPCR reaction products with the ssDNA-specific ExoI or S1 nucleases or dsDNA-specific restriction enzymes Eco RI and Nae I (left panel). Agarose gel electrophoresis of the digestion products as labeled by lane (right panel). M: Marker, C: aPCR product control, ExoI: exonuclease I, S1: S1 nuclease, Enz: Eco RI + Nae I. ( c ) NEB LongAmp was used to generate ssDNA up to 15,000 nt long using lambda phage dsDNA as template. Purification of the 10 kb fragment shows a single band of higher molecular weight than the M13mp18 ssDNA (7,249 nt). ( d ) The primer design algorithm aPrime was used to select primers for product sizes between 500 and 3,000 nt using M13mp18 ssDNA as template and the Quantabio Accustart HiFi enzyme. SYBR Safe stained agarose gels illuminated under blue light show dsDNA as yellow bands, while ssDNA show as orange bands.

    Journal: Scientific Reports

    Article Title: In vitro synthesis of gene-length single-stranded DNA

    doi: 10.1038/s41598-018-24677-5

    Figure Lengend Snippet: ssDNA production by aPCR. ( a ) aPCR reactions were assembled with a 50-molar excess of a forward primer for the amplification of a 1,000 nt ssDNA fragment using the M13mp18 ssDNA plasmid as template, and with 10 different polymerases that were tested for highest yield of ssDNA production (upper band: expected dsDNA size is 1,000 bp; lower band: expected ssDNA size is 1,000 nt) as judged by agarose gel electrophoresis (right panel). QuantaBio AccuStart HiFi, polymerase (lane 2, boxed) produced the highest amount without overlapping dsDNA contaminants. 1. Accustart; 2. Accustart HiFi; 3. Accustart II; 4. AccuPrime; 5. GoTaq; 6. DreamTaq; 7. Phusion; 8. Platinum SuperFi; 9. Q5; 10. Tth polymerase. ( b ) Biochemical validation of ssDNA production by incubating 1,000 nt aPCR reaction products with the ssDNA-specific ExoI or S1 nucleases or dsDNA-specific restriction enzymes Eco RI and Nae I (left panel). Agarose gel electrophoresis of the digestion products as labeled by lane (right panel). M: Marker, C: aPCR product control, ExoI: exonuclease I, S1: S1 nuclease, Enz: Eco RI + Nae I. ( c ) NEB LongAmp was used to generate ssDNA up to 15,000 nt long using lambda phage dsDNA as template. Purification of the 10 kb fragment shows a single band of higher molecular weight than the M13mp18 ssDNA (7,249 nt). ( d ) The primer design algorithm aPrime was used to select primers for product sizes between 500 and 3,000 nt using M13mp18 ssDNA as template and the Quantabio Accustart HiFi enzyme. SYBR Safe stained agarose gels illuminated under blue light show dsDNA as yellow bands, while ssDNA show as orange bands.

    Article Snippet: Initial tests with two other Taq -based polymerase sets, NEB LongAmp® Taq and Takara LA® Taq , produced notable amounts of dsDNA byproduct when tested for amplification of the 1,000 nt and the 3,281 nt fragments and reduced amount of ssDNA per reaction for the 1,000 and 3,281 fragments respectively in comparison with the Accustart HiFi (Fig. and Supplementary Table , External Table ).

    Techniques: Amplification, Plasmid Preparation, Agarose Gel Electrophoresis, Produced, Labeling, Marker, Purification, Molecular Weight, Staining