phusion hf dna polymerase new england biolabs  (New England Biolabs)


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    Name:
    Phusion High Fidelity DNA Polymerase
    Description:
    Phusion High Fidelity DNA Polymerase 500 units
    Catalog Number:
    m0530l
    Price:
    446
    Size:
    500 units
    Category:
    Thermostable DNA Polymerases
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    Structured Review

    New England Biolabs phusion hf dna polymerase new england biolabs
    Phusion High Fidelity DNA Polymerase
    Phusion High Fidelity DNA Polymerase 500 units
    https://www.bioz.com/result/phusion hf dna polymerase new england biolabs/product/New England Biolabs
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    phusion hf dna polymerase new england biolabs - by Bioz Stars, 2020-08
    99/100 stars

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    Related Articles

    Sequencing:

    Article Title: Deep Sequencing for Evaluation of Genetic Stability of Influenza A/California/07/2009 (H1N1) Vaccine Viruses
    Article Snippet: .. This result demonstrated that adequate distribution of sequencing reads between segments were obtained from a DNA library prepared from samples amplified by Phusion DNA polymerase (DNA library), and by whole-RNA library. .. Consistency of Illumina sequencing, and sequencing analysis of A/PR/8/34 and A/California/07/2009 strains Center for Biologics Evaluation and Research (CBER) stock of A/PR/8/34 was kindly provided by Dr. Peter Palese at Mount.

    Article Title: Demonstration of the Presence of the “Deleted” MIR122 Gene in HepG2 Cells
    Article Snippet: .. Such changes were still observed after amplification with Phusion High Fidelity DNA Polymerase, with 3/15 (20%) of single allele clones obtained from HepG2 and Huh-7 DNA showing apparent poly(T) slippage and also four sequence variants observed that were not seen in other clones of the same haplotype ( ). .. Overall, despite this sequence heterogeneity, the polymorphisms confirmed two different haplotypes in HepG2 DNA, consistent with the presence of two alleles of the pre-mir-122 stem-loop region.

    Clone Assay:

    Article Title: Demonstration of the Presence of the “Deleted” MIR122 Gene in HepG2 Cells
    Article Snippet: .. Such changes were still observed after amplification with Phusion High Fidelity DNA Polymerase, with 3/15 (20%) of single allele clones obtained from HepG2 and Huh-7 DNA showing apparent poly(T) slippage and also four sequence variants observed that were not seen in other clones of the same haplotype ( ). .. Overall, despite this sequence heterogeneity, the polymorphisms confirmed two different haplotypes in HepG2 DNA, consistent with the presence of two alleles of the pre-mir-122 stem-loop region.

    Amplification:

    Article Title: Deep Sequencing for Evaluation of Genetic Stability of Influenza A/California/07/2009 (H1N1) Vaccine Viruses
    Article Snippet: .. This result demonstrated that adequate distribution of sequencing reads between segments were obtained from a DNA library prepared from samples amplified by Phusion DNA polymerase (DNA library), and by whole-RNA library. .. Consistency of Illumina sequencing, and sequencing analysis of A/PR/8/34 and A/California/07/2009 strains Center for Biologics Evaluation and Research (CBER) stock of A/PR/8/34 was kindly provided by Dr. Peter Palese at Mount.

    Article Title: Demonstration of the Presence of the “Deleted” MIR122 Gene in HepG2 Cells
    Article Snippet: .. Such changes were still observed after amplification with Phusion High Fidelity DNA Polymerase, with 3/15 (20%) of single allele clones obtained from HepG2 and Huh-7 DNA showing apparent poly(T) slippage and also four sequence variants observed that were not seen in other clones of the same haplotype ( ). .. Overall, despite this sequence heterogeneity, the polymorphisms confirmed two different haplotypes in HepG2 DNA, consistent with the presence of two alleles of the pre-mir-122 stem-loop region.

    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
    Article Snippet: .. Successful PCR amplification has also been obtained with a Phusion High‐Fidelity DNA polymerase (New England Biolabs; M0530; see Fig. A). .. This polymerase was tested in the following PCR conditions for PCR products of ∼1.2 kb: 1 cycle at 98°C for 30 s followed by 35 cycles of 98°C for 10 s, 58°C for 20 s, 72°C for 45 s, and finally 1 cycle of 72°C for 5 min. 8 Following the PCR, mix 5 µl of the PCR reaction with 1 µl of 6× DNA loading dye and load the reactions on an agarose gel (Voytas, ).

    DNA Purification:

    Article Title: Enzymatic Synthesis of Modified Oligonucleotides by PEAR Using Phusion and KOD DNA Polymerases
    Article Snippet: .. Four 2′-fluoro-2′-deoxyribinucleoside-5′-triphosphates (2′-F-dNTPs), including 2′-F-dATP, 2′-F-dCTP, 2′-F-dGTP, 2′-F-dUTP and four 2′-deoxyribonucleotides-5′-O-(1-thiotriphosphate) (dNTPαSs), including dATPαS, dGTPαS, dCTPαS, and dTTPαS, whose structural formula are shown in , were purchased from Trilink BioTechnologies, Inc. KOD DNA polymerase was purchased from TOYOBO (Shanghai) Biotech Co., Ltd. Phusion DNA polymerase, highly thermostable restriction enzyme PspGI, and dNTPs were purchased from New England Biolabs, Inc. UNIQ-10 Spin Column Oligo DNA Purification Kit was purchased from Sangon Biotech (Shanghai) Co., Ltd. .. Synthetic oligodeoxynucleotides, including a target ( X ) and a probe ( P ), were synthesized by Integrated DNA Technologies, Inc. and purified by high-performance liquid chromatography (HPLC).

    Polymerase Chain Reaction:

    Article Title: Variations of five eIF4E genes across cassava accessions exhibiting tolerant and susceptible responses to cassava brown streak disease
    Article Snippet: .. PCR was performed in a 20 μl reaction volume containing 10 unit Phusion DNA polymerase (NEB, Ipswich, MA), 1 μl of 1:5 diluted cDNA template, 1X Phusion PCR buffer, 5 μM each of upstream and downstream primers, and 250 nM dNTP with the following cycling condition: 98°C for 1 minute; 35 cycles of 98°C for 15 seconds, 56°C for 15 seconds, and 72°C for 45 seconds; and finally 72°C for 5 minutes. .. Primers were designed according to five annotated eIF4E transcripts identified in the draft cassava genomic sequence (Manihot esculenta v4.1) published in Phytozome ( http://phytozome.jgi.doe.gov ) in 2013, prior to the availability of the current cassava genome V6.1 ( ).

    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
    Article Snippet: .. Successful PCR amplification has also been obtained with a Phusion High‐Fidelity DNA polymerase (New England Biolabs; M0530; see Fig. A). .. This polymerase was tested in the following PCR conditions for PCR products of ∼1.2 kb: 1 cycle at 98°C for 30 s followed by 35 cycles of 98°C for 10 s, 58°C for 20 s, 72°C for 45 s, and finally 1 cycle of 72°C for 5 min. 8 Following the PCR, mix 5 µl of the PCR reaction with 1 µl of 6× DNA loading dye and load the reactions on an agarose gel (Voytas, ).

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  • 99
    New England Biolabs phusion high fidelity pcr master mix with hf buffer
    Phusion High Fidelity Pcr Master Mix With Hf Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phusion high fidelity pcr master mix with hf buffer/product/New England Biolabs
    Average 99 stars, based on 77 article reviews
    Price from $9.99 to $1999.99
    phusion high fidelity pcr master mix with hf buffer - by Bioz Stars, 2020-08
    99/100 stars
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    99
    New England Biolabs yes pcr polymerase kit phusion high fidelity dna polymerase
    Screening sgRNAs for cleavage activity in vivo. ( A ) Schematic of the screening assay. Individual embryos are injected with RNPs composed of a particular sgRNA. Genomic <t>DNA</t> from each embryo is <t>PCR-amplified,</t> and amplicons are denatured and re-annealed. Heteroduplexes with mismatches due to indels in embryonic DNA are cleaved by T7E1 enzyme. Gel electrophoresis identifies embryos with detectable cleavage events. ( B ) PCR products of a target site in the forked gene 892 bp in length were digested by T7E1 as indicated. Shown are two representative embryos out of the nine assayed that were injected with forked RNPs. Also shown are two out of the six embryos that were uninjected. The predicted T7E1 digest products are 393 and 436 bp. Although a minority of heteroduplexes derived from an embryo are T7E1-sensitive, they can be detected by this assay. ( C ) A T7E1 assay performed on a sgRNA that was inactive in vivo. The target region is located in non-coding DNA. Three of the 12 RNP-injected embryo samples are shown, and three of the six uninjected embryo samples are shown. Heteroduplexes from the uninjected samples show T7E1 sensitivity that is likely due to sequence polymorphisms or non-B form DNA structures. The predicted T7E1 digest products from NHEJ induced mismatches are 295 and 502 bp. Note that samples from RNP-injected embryos do not exhibit T7E1 products of those sizes.
    Yes Pcr Polymerase Kit Phusion High Fidelity Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 3117 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/yes pcr polymerase kit phusion high fidelity dna polymerase/product/New England Biolabs
    Average 99 stars, based on 3117 article reviews
    Price from $9.99 to $1999.99
    yes pcr polymerase kit phusion high fidelity dna polymerase - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

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    Screening sgRNAs for cleavage activity in vivo. ( A ) Schematic of the screening assay. Individual embryos are injected with RNPs composed of a particular sgRNA. Genomic DNA from each embryo is PCR-amplified, and amplicons are denatured and re-annealed. Heteroduplexes with mismatches due to indels in embryonic DNA are cleaved by T7E1 enzyme. Gel electrophoresis identifies embryos with detectable cleavage events. ( B ) PCR products of a target site in the forked gene 892 bp in length were digested by T7E1 as indicated. Shown are two representative embryos out of the nine assayed that were injected with forked RNPs. Also shown are two out of the six embryos that were uninjected. The predicted T7E1 digest products are 393 and 436 bp. Although a minority of heteroduplexes derived from an embryo are T7E1-sensitive, they can be detected by this assay. ( C ) A T7E1 assay performed on a sgRNA that was inactive in vivo. The target region is located in non-coding DNA. Three of the 12 RNP-injected embryo samples are shown, and three of the six uninjected embryo samples are shown. Heteroduplexes from the uninjected samples show T7E1 sensitivity that is likely due to sequence polymorphisms or non-B form DNA structures. The predicted T7E1 digest products from NHEJ induced mismatches are 295 and 502 bp. Note that samples from RNP-injected embryos do not exhibit T7E1 products of those sizes.

    Journal: bioRxiv

    Article Title: Adaptable and Efficient Genome Editing by sgRNA-Cas9 Protein Co-injection into Drosophila

    doi: 10.1101/2020.05.07.080762

    Figure Lengend Snippet: Screening sgRNAs for cleavage activity in vivo. ( A ) Schematic of the screening assay. Individual embryos are injected with RNPs composed of a particular sgRNA. Genomic DNA from each embryo is PCR-amplified, and amplicons are denatured and re-annealed. Heteroduplexes with mismatches due to indels in embryonic DNA are cleaved by T7E1 enzyme. Gel electrophoresis identifies embryos with detectable cleavage events. ( B ) PCR products of a target site in the forked gene 892 bp in length were digested by T7E1 as indicated. Shown are two representative embryos out of the nine assayed that were injected with forked RNPs. Also shown are two out of the six embryos that were uninjected. The predicted T7E1 digest products are 393 and 436 bp. Although a minority of heteroduplexes derived from an embryo are T7E1-sensitive, they can be detected by this assay. ( C ) A T7E1 assay performed on a sgRNA that was inactive in vivo. The target region is located in non-coding DNA. Three of the 12 RNP-injected embryo samples are shown, and three of the six uninjected embryo samples are shown. Heteroduplexes from the uninjected samples show T7E1 sensitivity that is likely due to sequence polymorphisms or non-B form DNA structures. The predicted T7E1 digest products from NHEJ induced mismatches are 295 and 502 bp. Note that samples from RNP-injected embryos do not exhibit T7E1 products of those sizes.

    Article Snippet: To design ideal primers to generate DNA fragments for Gibson assembly, use the NEBuilder tool with the following build settings: http://nebuilder.neb.com/#!/ Product Kit: NEBuilder HiFi DNA Assembly Master Mix Minimum Overlap: 30 nt Circularize: Yes PCR Polymerase/Kit: Phusion High-Fidelity DNA Polymerase (HF Buffer) PCR Primer Conc.

    Techniques: Activity Assay, In Vivo, Screening Assay, Injection, Polymerase Chain Reaction, Amplification, Nucleic Acid Electrophoresis, Derivative Assay, Sequencing, Non-Homologous End Joining

    “Dissection” of the Gibson assembly into its component reactions. In the Gibson assembly, single-stranded 3’-overhangs are produced using T5 exonuclease. After insert-to-plasmid annealing at complementary single-stranded DNA ends, gaps are filled-in by Phusion DNA polymerase, and finally, Taq DNA ligase ligates the nicks. Here, we compared the efficiencies at which insert-plasmid mixtures transformed chemically competent E . coli cells. We used untreated insert-plasmid mixtures (co-transformation cloning), insert-plasmid mixtures treated with T5 exonuclease (sequence- and ligation-independent cloning), insert-plasmid mixtures treated with T5 exonuclease and Phusion DNA polymerase (sequence- and ligation-independent cloning plus gap filling), and insert-plasmid mixtures treated with T5 exonuclease, Phusion DNA polymerase and Taq DNA ligase (Gibson assembly). A) Data points represent the number of positive (blue) colonies averaged over three experiments ±SD. B) Data points represent the percentage of positive colonies averaged over three experiments ±SD.

    Journal: PLoS ONE

    Article Title: Seamless Insert-Plasmid Assembly at High Efficiency and Low Cost

    doi: 10.1371/journal.pone.0153158

    Figure Lengend Snippet: “Dissection” of the Gibson assembly into its component reactions. In the Gibson assembly, single-stranded 3’-overhangs are produced using T5 exonuclease. After insert-to-plasmid annealing at complementary single-stranded DNA ends, gaps are filled-in by Phusion DNA polymerase, and finally, Taq DNA ligase ligates the nicks. Here, we compared the efficiencies at which insert-plasmid mixtures transformed chemically competent E . coli cells. We used untreated insert-plasmid mixtures (co-transformation cloning), insert-plasmid mixtures treated with T5 exonuclease (sequence- and ligation-independent cloning), insert-plasmid mixtures treated with T5 exonuclease and Phusion DNA polymerase (sequence- and ligation-independent cloning plus gap filling), and insert-plasmid mixtures treated with T5 exonuclease, Phusion DNA polymerase and Taq DNA ligase (Gibson assembly). A) Data points represent the number of positive (blue) colonies averaged over three experiments ±SD. B) Data points represent the percentage of positive colonies averaged over three experiments ±SD.

    Article Snippet: Each Gibson assembly reaction consisted of 2.7 μl 5x IT buffer, 2 μl insert-plasmid mastermix (containing 75 ng plasmid and an 8-fold molar excess of insert), 5.3 μl 1:1000 diluted T5 exonuclease (New England Biolabs M0363S, 10’000 U/ml), 1.6 μl of 1:10 diluted Phusion HF DNA polymerase (NEB M0530L, 2’000 U/ml), 1.3 μl Taq DNA ligase (NEB M0208L, 40’000 U/ml, undiluted) and H2 0 to a final volume of 13.5 μl.

    Techniques: Produced, Plasmid Preparation, Transformation Assay, Clone Assay, Sequencing, Ligation