phusion polymerase  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99

    Structured Review

    New England Biolabs phusion polymerase
    Phusion Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 790 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phusion polymerase/product/New England Biolabs
    Average 99 stars, based on 790 article reviews
    Price from $9.99 to $1999.99
    phusion polymerase - by Bioz Stars, 2020-04
    99/100 stars

    Images

    Related Articles

    Clone Assay:

    Article Title: CRISPR–Cas9-enabled genetic disruptions for understanding ethanol and ethyl acetate biosynthesis in Kluyveromyces marxianus
    Article Snippet: .. Molecular cloning and plasmids construction All cloning was accomplished using Phusion polymerase, restriction endonucleases, and Gibson assembly master mix purchased from New England BioLabs (NEB). .. DNA oligos were purchased from Integrated DNA Technologies (IDT).

    Article Title: A Chemical Genetic Screen Identifies ABHD12 as an Oxidized Phosphatidylserine Lipase
    Article Snippet: Expression of recombinant human ABHD12 The full-length hABHD12 cDNA was cloned into pCMV-Sport6 vector between NotI and SalI restriction sites (GE Life Sciences). .. The S246A hABHD12 mutant was generated from the same plasmid using Phusion polymerase and DpnI (New England Biolabs) as per manufacturers instructions.

    Article Title: Structures of β-glycosidase LXYL-P1-2 reveals the product binding state of GH3 family and a specific pocket for Taxol recognition
    Article Snippet: Materials and strains The plasmid pPIC3.5K-lxyl-p1 - 2 was cloned in our lab , . .. Phusion polymerase, restriction enzymes, and T4 ligase were purchased from New England Biolabs (Ipswich, MA).

    Amplification:

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: .. Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material. .. For the Hi-C sample, preparation was stopped here and the sample corresponding to material treated with Interferonγ sequenced.

    Synthesized:

    Article Title: Structures of β-glycosidase LXYL-P1-2 reveals the product binding state of GH3 family and a specific pocket for Taxol recognition
    Article Snippet: The TnBgl3B gene (GenBank: ABI29899.1) was synthesized by SynBio Research Platform at Tianjin University (Tianjin, China). .. Phusion polymerase, restriction enzymes, and T4 ligase were purchased from New England Biolabs (Ipswich, MA).

    Incubation:

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material. .. Briefly, 1 μg of Hi-C material was incubated at 42 °C for 48 h with 2.89 μmol of a MER41 biotinylated DNA oligo according to Nimblegen’s instructions.

    Activity Assay:

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: A-tailing was done using Klenow polymerase without 3′ exonuclease activity. .. Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material.

    Expressing:

    Article Title: A Chemical Genetic Screen Identifies ABHD12 as an Oxidized Phosphatidylserine Lipase
    Article Snippet: Paragraph title: Expression of recombinant human ABHD12 ... The S246A hABHD12 mutant was generated from the same plasmid using Phusion polymerase and DpnI (New England Biolabs) as per manufacturers instructions.

    Transformation Assay:

    Article Title: Restraint of the G2/M Transition by the SR/RRM Family mRNA Shuttling Binding Protein SNXAHRB1 in Aspergillus nidulans
    Article Snippet: Phusion Polymerase was used for all PCR experiments, and all other DNA-modifying enzymes were from New England BioLabs. .. Standard methods of Aspergillus culture , genetic analysis ( ; ), construction and analysis of heterokaryons and diploids , and Aspergillus transformation ( ) were employed.

    Kinase Assay:

    Article Title: Restraint of the G2/M Transition by the SR/RRM Family mRNA Shuttling Binding Protein SNXAHRB1 in Aspergillus nidulans
    Article Snippet: Phusion Polymerase was used for all PCR experiments, and all other DNA-modifying enzymes were from New England BioLabs. .. Rich media, composed of minimal medium ( ) supplemented with 0.5% yeast extract and containing 2% glucose, were used for kinase assay experiments and to grow cells for DNA-mediated transformation.

    Hybridization:

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material. .. For Capture, we used the Nimblegen SeqCap EZ hybridization and wash kit (05634261001), Nimblegen SeqCap EZ accessory kit v2 and Nimblegen SeqCap EZ HE-oligo kit.

    High Performance Liquid Chromatography:

    Article Title: Structures of β-glycosidase LXYL-P1-2 reveals the product binding state of GH3 family and a specific pocket for Taxol recognition
    Article Snippet: Phusion polymerase, restriction enzymes, and T4 ligase were purchased from New England Biolabs (Ipswich, MA). .. XDT and DT (HPLC purity > 99%) were purified in our laboratory.

    Transfection:

    Article Title: A Chemical Genetic Screen Identifies ABHD12 as an Oxidized Phosphatidylserine Lipase
    Article Snippet: The S246A hABHD12 mutant was generated from the same plasmid using Phusion polymerase and DpnI (New England Biolabs) as per manufacturers instructions. .. Recombinant WT and S246A hABHD12 was generated from HEK293T cells using established transient transfection protocols .

    Inverse PCR:

    Article Title: Tcrd rearrangement redirects a processive Tcra recombination program to expand the Tcra repertoire
    Article Snippet: Each library was then used for two rounds of inverse PCR from each of four viewpoints, with eight individual PCR reactions performed per viewpoint. .. All PCR reactions used Phusion polymerase in Phusion HiFi buffer (New England Biolabs) at a reaction volume of 50 μl.

    Introduce:

    Article Title: Control of septin filament flexibility and bundling by subunit composition and nucleotide interactions
    Article Snippet: Custom oligonucleotides were from Integrated DNA Technologies (IDT, Coralville, IA), and Phusion polymerase was used for PCR (NEB, Ipswitch, MA). .. To generate plasmid pMVB128 HIS-Sccdc12 T75A/ScCDC10 (AGB561), AGO1340 and AGO1341 were used to introduce the point mutation using the Agilent XL QuickChange kit.

    Generated:

    Article Title: A Chemical Genetic Screen Identifies ABHD12 as an Oxidized Phosphatidylserine Lipase
    Article Snippet: .. The S246A hABHD12 mutant was generated from the same plasmid using Phusion polymerase and DpnI (New England Biolabs) as per manufacturers instructions. .. Recombinant WT and S246A hABHD12 was generated from HEK293T cells using established transient transfection protocols .

    Article Title: Ancestral population reconstitution from isofemale lines as a tool for experimental evolution
    Article Snippet: For the D. melanogaster RAPs, libraries were generated from 4 μg genomic DNA again using NEBNext Mastermix reagents followed by bead‐based size selection for an insert size of 330 bp. .. PCR was carried out using Phusion polymerase with six cycles at 65°C and NEBNext single index barcodes.

    DNA Sequencing:

    Article Title: A broadly distributed toxin family mediates contact-dependent antagonism between gram-positive bacteria
    Article Snippet: Phusion polymerase, restriction enzymes and T4 DNA ligase were obtained from New England Biolabs (NEB). .. Phusion polymerase, restriction enzymes and T4 DNA ligase were obtained from New England Biolabs (NEB).

    Article Title: EspH is a hypervirulence factor for Mycobacterium marinum and essential for the secretion of the ESX-1 substrates EspE and EspF
    Article Snippet: Phusion polymerase, restriction enzymes and T4 DNA ligase were obtained from New England Biolabs (NEB). .. Macrogen performed DNA sequencing.

    Sequencing:

    Article Title: Control of septin filament flexibility and bundling by subunit composition and nucleotide interactions
    Article Snippet: Custom oligonucleotides were from Integrated DNA Technologies (IDT, Coralville, IA), and Phusion polymerase was used for PCR (NEB, Ipswitch, MA). .. The correct product was verified using digestion with Eco RV and sequencing using AGO1192 and AGO1342.

    Article Title: Ancestral population reconstitution from isofemale lines as a tool for experimental evolution
    Article Snippet: Paragraph title: Genome sequencing ... PCR was carried out using Phusion polymerase with six cycles at 65°C and NEBNext single index barcodes.

    Article Title: Tcrd rearrangement redirects a processive Tcra recombination program to expand the Tcra repertoire
    Article Snippet: PCR conditions were as follows: 30 s at 98°C, followed by 20 cycles of 10 s at 98°C, 30 s at 60°C and 2 min at 72°C, with a final extension for 10 min at 72°C PCR products were purified with QiaQuick PCR purification reagents (Qiagen) and UPrep spin columns (Genesee), and were then subjected to second round PCR to add Illumina sequencing adapters to their ends. .. All PCR reactions used Phusion polymerase in Phusion HiFi buffer (New England Biolabs) at a reaction volume of 50 μl.

    Sonication:

    Article Title: A Chemical Genetic Screen Identifies ABHD12 as an Oxidized Phosphatidylserine Lipase
    Article Snippet: The S246A hABHD12 mutant was generated from the same plasmid using Phusion polymerase and DpnI (New England Biolabs) as per manufacturers instructions. .. The cells were harvested by scraping 48 h after transfection, washed with sterile DPBS (3X), resuspended in 1 mL DPBS, and lysed by sonication.

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: DNA fragments were then bound to Streptavidin C1 beads and washed following manufacturer’s instructions and using 150 μl per 2.5 μg of sonicated DNA. .. Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material.

    Binding Assay:

    Article Title: Trichophyton rubrum LysM proteins bind to fungal cell wall chitin and to the N-linked oligosaccharides present on human skin glycoproteins
    Article Snippet: Amylose resins, chitin magnetic beads, restriction enzymes, Phusion polymerase, PNGase F, and T4 DNA ligase were purchased from New England Biolabs (Ipswich, MA). .. Antibody directed against Maltose Binding Protein (product E8032) was obtained from New England Biolabs and used according to the manufacturer’s instructions.

    Hi-C:

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: .. Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material. .. For the Hi-C sample, preparation was stopped here and the sample corresponding to material treated with Interferonγ sequenced.

    Nucleic Acid Electrophoresis:

    Article Title: Tcrd rearrangement redirects a processive Tcra recombination program to expand the Tcra repertoire
    Article Snippet: All PCR reactions used Phusion polymerase in Phusion HiFi buffer (New England Biolabs) at a reaction volume of 50 μl. .. Following second round PCR, the eight reactions for each viewpoint were pooled, and PCR products were purified as described above and assessed by gel electrophoresis.

    Magnetic Beads:

    Article Title: Trichophyton rubrum LysM proteins bind to fungal cell wall chitin and to the N-linked oligosaccharides present on human skin glycoproteins
    Article Snippet: .. Amylose resins, chitin magnetic beads, restriction enzymes, Phusion polymerase, PNGase F, and T4 DNA ligase were purchased from New England Biolabs (Ipswich, MA). .. Antibody directed against Maltose Binding Protein (product E8032) was obtained from New England Biolabs and used according to the manufacturer’s instructions.

    Mutagenesis:

    Article Title: A Chemical Genetic Screen Identifies ABHD12 as an Oxidized Phosphatidylserine Lipase
    Article Snippet: .. The S246A hABHD12 mutant was generated from the same plasmid using Phusion polymerase and DpnI (New England Biolabs) as per manufacturers instructions. .. Recombinant WT and S246A hABHD12 was generated from HEK293T cells using established transient transfection protocols .

    Article Title: Control of septin filament flexibility and bundling by subunit composition and nucleotide interactions
    Article Snippet: Custom oligonucleotides were from Integrated DNA Technologies (IDT, Coralville, IA), and Phusion polymerase was used for PCR (NEB, Ipswitch, MA). .. To generate plasmid pMVB128 HIS-Sccdc12 T75A/ScCDC10 (AGB561), AGO1340 and AGO1341 were used to introduce the point mutation using the Agilent XL QuickChange kit.

    Purification:

    Article Title: Structures of β-glycosidase LXYL-P1-2 reveals the product binding state of GH3 family and a specific pocket for Taxol recognition
    Article Snippet: Phusion polymerase, restriction enzymes, and T4 ligase were purchased from New England Biolabs (Ipswich, MA). .. XDT and DT (HPLC purity > 99%) were purified in our laboratory.

    Article Title: Tcrd rearrangement redirects a processive Tcra recombination program to expand the Tcra repertoire
    Article Snippet: PCR conditions were as follows: 30 s at 98°C, followed by 20 cycles of 10 s at 98°C, 30 s at 60°C and 2 min at 72°C, with a final extension for 10 min at 72°C PCR products were purified with QiaQuick PCR purification reagents (Qiagen) and UPrep spin columns (Genesee), and were then subjected to second round PCR to add Illumina sequencing adapters to their ends. .. All PCR reactions used Phusion polymerase in Phusion HiFi buffer (New England Biolabs) at a reaction volume of 50 μl.

    Polymerase Chain Reaction:

    Article Title: Restraint of the G2/M Transition by the SR/RRM Family mRNA Shuttling Binding Protein SNXAHRB1 in Aspergillus nidulans
    Article Snippet: .. Phusion Polymerase was used for all PCR experiments, and all other DNA-modifying enzymes were from New England BioLabs. .. Standard methods of Aspergillus culture , genetic analysis ( ; ), construction and analysis of heterokaryons and diploids , and Aspergillus transformation ( ) were employed.

    Article Title: Control of septin filament flexibility and bundling by subunit composition and nucleotide interactions
    Article Snippet: .. Custom oligonucleotides were from Integrated DNA Technologies (IDT, Coralville, IA), and Phusion polymerase was used for PCR (NEB, Ipswitch, MA). .. To generate plasmid pMVB128 HIS-Sccdc12 T75A/ScCDC10 (AGB561), AGO1340 and AGO1341 were used to introduce the point mutation using the Agilent XL QuickChange kit.

    Article Title: Ancestral population reconstitution from isofemale lines as a tool for experimental evolution
    Article Snippet: .. PCR was carried out using Phusion polymerase with six cycles at 65°C and NEBNext single index barcodes. .. Libraries for the D. simulans APs were prepared from 2.5 μg genomic DNA using the TruSeq DNA PCR‐free library preparation protocol (Illumina Inc., San Diego, CA) with size selection for 420‐bp fragment size and single index adapters.

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: .. Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material. .. For the Hi-C sample, preparation was stopped here and the sample corresponding to material treated with Interferonγ sequenced.

    Article Title: Tcrd rearrangement redirects a processive Tcra recombination program to expand the Tcra repertoire
    Article Snippet: .. All PCR reactions used Phusion polymerase in Phusion HiFi buffer (New England Biolabs) at a reaction volume of 50 μl. .. Following second round PCR, the eight reactions for each viewpoint were pooled, and PCR products were purified as described above and assessed by gel electrophoresis.

    Selection:

    Article Title: Ancestral population reconstitution from isofemale lines as a tool for experimental evolution
    Article Snippet: For the D. melanogaster RAPs, libraries were generated from 4 μg genomic DNA again using NEBNext Mastermix reagents followed by bead‐based size selection for an insert size of 330 bp. .. PCR was carried out using Phusion polymerase with six cycles at 65°C and NEBNext single index barcodes.

    Agarose Gel Electrophoresis:

    Article Title: Ancestral population reconstitution from isofemale lines as a tool for experimental evolution
    Article Snippet: The insert size distribution of the final libraries was tightened to a mean of approximately 280 bp by size selection on an agarose gel. .. PCR was carried out using Phusion polymerase with six cycles at 65°C and NEBNext single index barcodes.

    Plasmid Preparation:

    Article Title: CRISPR–Cas9-enabled genetic disruptions for understanding ethanol and ethyl acetate biosynthesis in Kluyveromyces marxianus
    Article Snippet: Molecular cloning and plasmids construction All cloning was accomplished using Phusion polymerase, restriction endonucleases, and Gibson assembly master mix purchased from New England BioLabs (NEB). .. Chemically competent DH5α Escherichia coli was used for plasmid propagation.

    Article Title: A Chemical Genetic Screen Identifies ABHD12 as an Oxidized Phosphatidylserine Lipase
    Article Snippet: .. The S246A hABHD12 mutant was generated from the same plasmid using Phusion polymerase and DpnI (New England Biolabs) as per manufacturers instructions. .. Recombinant WT and S246A hABHD12 was generated from HEK293T cells using established transient transfection protocols .

    Article Title: Control of septin filament flexibility and bundling by subunit composition and nucleotide interactions
    Article Snippet: Custom oligonucleotides were from Integrated DNA Technologies (IDT, Coralville, IA), and Phusion polymerase was used for PCR (NEB, Ipswitch, MA). .. To generate plasmid pMVB128 HIS-Sccdc12 T75A/ScCDC10 (AGB561), AGO1340 and AGO1341 were used to introduce the point mutation using the Agilent XL QuickChange kit.

    Article Title: A broadly distributed toxin family mediates contact-dependent antagonism between gram-positive bacteria
    Article Snippet: Paragraph title: DNA manipulation and plasmid construction ... Phusion polymerase, restriction enzymes and T4 DNA ligase were obtained from New England Biolabs (NEB).

    Article Title: EspH is a hypervirulence factor for Mycobacterium marinum and essential for the secretion of the ESX-1 substrates EspE and EspF
    Article Snippet: Paragraph title: DNA manipulation and plasmid construction ... Phusion polymerase, restriction enzymes and T4 DNA ligase were obtained from New England Biolabs (NEB).

    Article Title: Structures of β-glycosidase LXYL-P1-2 reveals the product binding state of GH3 family and a specific pocket for Taxol recognition
    Article Snippet: Materials and strains The plasmid pPIC3.5K-lxyl-p1 - 2 was cloned in our lab , . .. Phusion polymerase, restriction enzymes, and T4 ligase were purchased from New England Biolabs (Ipswich, MA).

    Multiplex Assay:

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: .. Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material. .. For the Hi-C sample, preparation was stopped here and the sample corresponding to material treated with Interferonγ sequenced.

    Positron Emission Tomography:

    Article Title: Structures of β-glycosidase LXYL-P1-2 reveals the product binding state of GH3 family and a specific pocket for Taxol recognition
    Article Snippet: Phusion polymerase, restriction enzymes, and T4 ligase were purchased from New England Biolabs (Ipswich, MA). .. The pET-28a plasmid was purchased from Novagen (Malaysia).

    Sample Prep:

    Article Title: Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate
    Article Snippet: Hi-C library amplification was then amplified using 7 PCR cycles, with Phusion polymerase and PCR primers from NEBNext Multiplex Oligos for Illumina (E7335) and resulted in approximately 1 μg of Hi-C material. .. For the Hi-C sample, preparation was stopped here and the sample corresponding to material treated with Interferonγ sequenced.

    Recombinant:

    Article Title: A Chemical Genetic Screen Identifies ABHD12 as an Oxidized Phosphatidylserine Lipase
    Article Snippet: Paragraph title: Expression of recombinant human ABHD12 ... The S246A hABHD12 mutant was generated from the same plasmid using Phusion polymerase and DpnI (New England Biolabs) as per manufacturers instructions.

    Molecular Cloning:

    Article Title: CRISPR–Cas9-enabled genetic disruptions for understanding ethanol and ethyl acetate biosynthesis in Kluyveromyces marxianus
    Article Snippet: .. Molecular cloning and plasmids construction All cloning was accomplished using Phusion polymerase, restriction endonucleases, and Gibson assembly master mix purchased from New England BioLabs (NEB). .. DNA oligos were purchased from Integrated DNA Technologies (IDT).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs phusion buffer hf
    Agarose gel electrophoresis analysis of PCR fragments multiplied by <t>Phusion</t> High-Fidelity PCR Master Mix with HF Buffer and Phusion High-Fidelity PCR Master Mix with GC Buffer. Analysed PCRs are labelled as the mutated residue and the letter indicating whether the fragment is multiplied by mutation-specific forward (F) or reverse (R) primer. Expected PCR fragments are between 3000 and 4000 bp long. While using the PCR master mix with GC buffer gave expected fragments in all shown cases, using the Phusion High-Fidelity polymerase with HF buffer failed to multiply four fragments.
    Phusion Buffer Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phusion buffer hf/product/New England Biolabs
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    phusion buffer hf - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    91
    New England Biolabs phusion hf pcr reactions
    Schematics of the subcycling <t>PCR</t> protocol. A. Each PCR cycle involves a denaturation step and an annealing/elongation step. We introduced 4X sub-cycling the annealing/elongation step within each of the 30X amplification cycles. B. Multiplexed amplification products for pools of 7 oligos with 154–200 bp length products of varying GC content. <t>Phusion</t> and KAPA HIFI polymerases were used with and without a sub-cycling thermocycle. Each different condition is used to amplify 12 separate oligo pools with GC content ranges as follows: 1.) 16.4–34.3; 2.) 13.5–38.7; 3.) 21.5–37.3; 4.) 12.2–12.2; 5.) 12.7–40.0; 6.) 14.9–41.6; 7.) 16.4–37.6; 8.) 12.7–42.0; 9.) 20.9–40.6; 10.) 12.5–42.5; 11.) 12.7–43.7; 12.) 14.8–35.6. Results are based on electronic gels created by electrophoresis using a Perkin Elmer GX instrument with a 5k chip. *Bin shows an example of an expected PCR pattern where a strong 154-200bp product band is seen. PCR reactions were not purified and primers can be seen at the bottom of each sample.
    Phusion Hf Pcr Reactions, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phusion hf pcr reactions/product/New England Biolabs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phusion hf pcr reactions - by Bioz Stars, 2020-04
    91/100 stars
      Buy from Supplier

    99
    New England Biolabs phusion high fidelity dna polymerase
    The median CEL intensities for each amplicon obtained by using Stoffel <t>DNA</t> polymerase and <t>Phusion</t> DNA polymerase in the gap-fill reaction are plotted against each other. The CEL intensities that were
    Phusion High Fidelity Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1359 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phusion high fidelity dna polymerase/product/New England Biolabs
    Average 99 stars, based on 1359 article reviews
    Price from $9.99 to $1999.99
    phusion high fidelity dna polymerase - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    Image Search Results


    Agarose gel electrophoresis analysis of PCR fragments multiplied by Phusion High-Fidelity PCR Master Mix with HF Buffer and Phusion High-Fidelity PCR Master Mix with GC Buffer. Analysed PCRs are labelled as the mutated residue and the letter indicating whether the fragment is multiplied by mutation-specific forward (F) or reverse (R) primer. Expected PCR fragments are between 3000 and 4000 bp long. While using the PCR master mix with GC buffer gave expected fragments in all shown cases, using the Phusion High-Fidelity polymerase with HF buffer failed to multiply four fragments.

    Journal: Scientific Reports

    Article Title: High-throughput mutagenesis using a two-fragment PCR approach

    doi: 10.1038/s41598-017-07010-4

    Figure Lengend Snippet: Agarose gel electrophoresis analysis of PCR fragments multiplied by Phusion High-Fidelity PCR Master Mix with HF Buffer and Phusion High-Fidelity PCR Master Mix with GC Buffer. Analysed PCRs are labelled as the mutated residue and the letter indicating whether the fragment is multiplied by mutation-specific forward (F) or reverse (R) primer. Expected PCR fragments are between 3000 and 4000 bp long. While using the PCR master mix with GC buffer gave expected fragments in all shown cases, using the Phusion High-Fidelity polymerase with HF buffer failed to multiply four fragments.

    Article Snippet: For truncations of the AtETR1 and LeETR1 plasmids by the two-fragment approach, a slightly different PCR protocol was used: 20 μL PCR mixture prepared with nuclease-free water (Cell Signaling Technology) contained 1 ng DNA template, 500 nM each primer, 200 μM each dNTP (deoxynucleoside triphosphate; NEB), 0.4 U μL−1 Phusion High-Fidelity DNA Polymerase (NEB), and either 1× Phusion buffer HF (NEB) or 3% (v/v) DMSO (NEB) with 1× Phusion buffer GC (NEB).

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Mutagenesis

    Schematics of the subcycling PCR protocol. A. Each PCR cycle involves a denaturation step and an annealing/elongation step. We introduced 4X sub-cycling the annealing/elongation step within each of the 30X amplification cycles. B. Multiplexed amplification products for pools of 7 oligos with 154–200 bp length products of varying GC content. Phusion and KAPA HIFI polymerases were used with and without a sub-cycling thermocycle. Each different condition is used to amplify 12 separate oligo pools with GC content ranges as follows: 1.) 16.4–34.3; 2.) 13.5–38.7; 3.) 21.5–37.3; 4.) 12.2–12.2; 5.) 12.7–40.0; 6.) 14.9–41.6; 7.) 16.4–37.6; 8.) 12.7–42.0; 9.) 20.9–40.6; 10.) 12.5–42.5; 11.) 12.7–43.7; 12.) 14.8–35.6. Results are based on electronic gels created by electrophoresis using a Perkin Elmer GX instrument with a 5k chip. *Bin shows an example of an expected PCR pattern where a strong 154-200bp product band is seen. PCR reactions were not purified and primers can be seen at the bottom of each sample.

    Journal: PLoS ONE

    Article Title: Improved PCR Amplification of Broad Spectrum GC DNA Templates

    doi: 10.1371/journal.pone.0156478

    Figure Lengend Snippet: Schematics of the subcycling PCR protocol. A. Each PCR cycle involves a denaturation step and an annealing/elongation step. We introduced 4X sub-cycling the annealing/elongation step within each of the 30X amplification cycles. B. Multiplexed amplification products for pools of 7 oligos with 154–200 bp length products of varying GC content. Phusion and KAPA HIFI polymerases were used with and without a sub-cycling thermocycle. Each different condition is used to amplify 12 separate oligo pools with GC content ranges as follows: 1.) 16.4–34.3; 2.) 13.5–38.7; 3.) 21.5–37.3; 4.) 12.2–12.2; 5.) 12.7–40.0; 6.) 14.9–41.6; 7.) 16.4–37.6; 8.) 12.7–42.0; 9.) 20.9–40.6; 10.) 12.5–42.5; 11.) 12.7–43.7; 12.) 14.8–35.6. Results are based on electronic gels created by electrophoresis using a Perkin Elmer GX instrument with a 5k chip. *Bin shows an example of an expected PCR pattern where a strong 154-200bp product band is seen. PCR reactions were not purified and primers can be seen at the bottom of each sample.

    Article Snippet: To address high GC content various additives were included in the Phusion HF PCR reactions as follows: 7-deaza-dGTP (NEB) at a 40:60 ratio with normal dGTP, as well as 50:50 and 60:40 ratios keeping the final concentration of dNTPs constant; DMSO (Sigma) at a final concentration of 2.5%, 5%, and 10%; betaine (Sigma) at a final concentration of 1M, 2M and 4M.

    Techniques: Polymerase Chain Reaction, Amplification, Electrophoresis, Chromatin Immunoprecipitation, Purification

    The median CEL intensities for each amplicon obtained by using Stoffel DNA polymerase and Phusion DNA polymerase in the gap-fill reaction are plotted against each other. The CEL intensities that were

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: A comprehensive assay for targeted multiplex amplification of human DNA sequences

    doi: 10.1073/pnas.0803240105

    Figure Lengend Snippet: The median CEL intensities for each amplicon obtained by using Stoffel DNA polymerase and Phusion DNA polymerase in the gap-fill reaction are plotted against each other. The CEL intensities that were

    Article Snippet: The extension was performed by addition of 0.4 units of Phusion High-Fidelity DNA Polymerase (New England Biolabs), 3 μl 1.0 mM dNTP, 5 units Ampligase (Epicenter Biotechnologies) in a 15-μl volume at 60°C for 15 min followed by 72°C for 15 min.

    Techniques: Amplification

    Polyacrylamide gel electrophoresis (PAGE) electrophoresis of the polymerase–endonuclease amplification reaction (PEAR) products. Lowercase letters (agct) represents unmodified dNTPs; uppercase letters (AGCT) represent modified dNTPs (2′-F-dNTPs or dNTPαSs). (A) PEAR by Phusion DNA polymerase using unmodified dNTPs, 2′-F-modified dATP and dGTP, respectively. Lane 1: 10-bp DNA ladder; lane 2: normal dNTPs; lane 3: 2′-F-dATP modified PEAR products; lane 4: control without PspGI; lane 5: control without Phusion DNA polymerase; lane 6: control without dATP; lane 7: 10-bp DNA ladder; lane 8: 2′-F-dGTP modified PEAR products; lane 9: control without PspGI; lane 10 : control without Phusion DNA polymerase; lane 11: control without dGTP. (B) PEAR by Phusion DNA polymerase using 2′-F-dCTP and 2′-F-dUTP. Lane 1: 2′-F-dCTP modified PEAR products; lane 2: control without PspGI; lane 3: control without Phusion DNA polymerase; lane 4: control without dCTP; lane 5: 10-bp DNA ladder; lane 6: 2′-F-dUTP modified PEAR products; lane 7: control without PspGI; lane 8: control without Phusion DNA polymerase; lane 9: control without dUTP; lane 10: 10-bp DNA ladder. (C) 2′-F-dATP and 2′-F-dGTP modified PEAR products as “seeds” for PEAR. Lane 1: 10-bp DNA ladder; lane 2: control without PspGI; lane 3: using 2′-F-dATP modified PEAR products as “seeds” for PEAR; lane 4: control without PspGI; lane 5: using 2′-F-dGTP modified PEAR products as seeds for PEAR. (D) 2′-F-dATP and 2′-F-dGTP modified PEAR products using KOD DNA polymerase. Lane 1: 2′-F-dATP modified PEAR products; lane 2: control without PspGI; lane 3: control without KOD DNA polymerase; lane 4: control without 2′-F-dATP; lane 5: 10-bp DNA ladder; lane 6: 2′-F-dGTP modified PEAR products; lane 7: control without PspGI; lane 8: control without KOD DNA polymerase ; lane 9: control without 2′-F-dGTP; lane 10: 10-bp DNA ladder. (E) PEAR amplification of 2′-F-dCTP and 2′-F-dUTP modified products using KOD DNA polymerase. Lane 1: 2′-F-dCTP modified PEAR products; lane 2: control without PspGI; lane 3: control without KOD DNA polymerase; lane 4: control without dCTP; lane 5: 10-bp DNA ladder; lane 6: 2′-F-dUTP modified PEAR products; lane 7: control without PspGI; lane 8: control without KOD DNA polymerase; lane 9: control without dUTP; lane 10: 10-bp DNA ladder. (F) PEAR amplification of dTTPαS modified and 2′-F-dATP+dGTPαS double modified PEAR products using KOD DNA polymerase. Lane 1: dTTPαS modified PEAR products; lane 2: control without PspGI; lane 3: control without KOD DNA polymerase; lane 4: control without dTTPαS; lane 5: 20 bp DNA ladder; lane 6: 2′-F-dATP and dGTPαS double modified PEAR amplified products; lane 7: control without PspGI; lane 8: control without KOD DNA polymerase; lane 9: control without 2′-F-dATP and dGTPαS; lane 10: 20-bp DNA ladder. (G) PEAR amplification of 2′-F-dATP+dCTPαS double modified and 2′-F-dATP+dTTPαS double modified PEAR products using KOD DNA polymerase. Lane 1: 2′-F-dATP+dCTPαS double modified PEAR products; lane 2: control without PspGI; lane 3: control without KOD DNA polymerase; lane 4: control without 2′-F-dATP and dCTPαS; lane 5: 20-bp DNA ladder; lane 6: 2′-F-dATP+dTTPαS double modified PEAR amplified products; lane 7: control without PspGI; lane 8: control without KOD DNA polymerase; lane 9: control without 2′-F-dGTP and dTTPαS; lane 10: 20-bp DNA ladder.

    Journal: Nucleic Acid Therapeutics

    Article Title: Enzymatic Synthesis of Modified Oligonucleotides by PEAR Using Phusion and KOD DNA Polymerases

    doi: 10.1089/nat.2014.0513

    Figure Lengend Snippet: Polyacrylamide gel electrophoresis (PAGE) electrophoresis of the polymerase–endonuclease amplification reaction (PEAR) products. Lowercase letters (agct) represents unmodified dNTPs; uppercase letters (AGCT) represent modified dNTPs (2′-F-dNTPs or dNTPαSs). (A) PEAR by Phusion DNA polymerase using unmodified dNTPs, 2′-F-modified dATP and dGTP, respectively. Lane 1: 10-bp DNA ladder; lane 2: normal dNTPs; lane 3: 2′-F-dATP modified PEAR products; lane 4: control without PspGI; lane 5: control without Phusion DNA polymerase; lane 6: control without dATP; lane 7: 10-bp DNA ladder; lane 8: 2′-F-dGTP modified PEAR products; lane 9: control without PspGI; lane 10 : control without Phusion DNA polymerase; lane 11: control without dGTP. (B) PEAR by Phusion DNA polymerase using 2′-F-dCTP and 2′-F-dUTP. Lane 1: 2′-F-dCTP modified PEAR products; lane 2: control without PspGI; lane 3: control without Phusion DNA polymerase; lane 4: control without dCTP; lane 5: 10-bp DNA ladder; lane 6: 2′-F-dUTP modified PEAR products; lane 7: control without PspGI; lane 8: control without Phusion DNA polymerase; lane 9: control without dUTP; lane 10: 10-bp DNA ladder. (C) 2′-F-dATP and 2′-F-dGTP modified PEAR products as “seeds” for PEAR. Lane 1: 10-bp DNA ladder; lane 2: control without PspGI; lane 3: using 2′-F-dATP modified PEAR products as “seeds” for PEAR; lane 4: control without PspGI; lane 5: using 2′-F-dGTP modified PEAR products as seeds for PEAR. (D) 2′-F-dATP and 2′-F-dGTP modified PEAR products using KOD DNA polymerase. Lane 1: 2′-F-dATP modified PEAR products; lane 2: control without PspGI; lane 3: control without KOD DNA polymerase; lane 4: control without 2′-F-dATP; lane 5: 10-bp DNA ladder; lane 6: 2′-F-dGTP modified PEAR products; lane 7: control without PspGI; lane 8: control without KOD DNA polymerase ; lane 9: control without 2′-F-dGTP; lane 10: 10-bp DNA ladder. (E) PEAR amplification of 2′-F-dCTP and 2′-F-dUTP modified products using KOD DNA polymerase. Lane 1: 2′-F-dCTP modified PEAR products; lane 2: control without PspGI; lane 3: control without KOD DNA polymerase; lane 4: control without dCTP; lane 5: 10-bp DNA ladder; lane 6: 2′-F-dUTP modified PEAR products; lane 7: control without PspGI; lane 8: control without KOD DNA polymerase; lane 9: control without dUTP; lane 10: 10-bp DNA ladder. (F) PEAR amplification of dTTPαS modified and 2′-F-dATP+dGTPαS double modified PEAR products using KOD DNA polymerase. Lane 1: dTTPαS modified PEAR products; lane 2: control without PspGI; lane 3: control without KOD DNA polymerase; lane 4: control without dTTPαS; lane 5: 20 bp DNA ladder; lane 6: 2′-F-dATP and dGTPαS double modified PEAR amplified products; lane 7: control without PspGI; lane 8: control without KOD DNA polymerase; lane 9: control without 2′-F-dATP and dGTPαS; lane 10: 20-bp DNA ladder. (G) PEAR amplification of 2′-F-dATP+dCTPαS double modified and 2′-F-dATP+dTTPαS double modified PEAR products using KOD DNA polymerase. Lane 1: 2′-F-dATP+dCTPαS double modified PEAR products; lane 2: control without PspGI; lane 3: control without KOD DNA polymerase; lane 4: control without 2′-F-dATP and dCTPαS; lane 5: 20-bp DNA ladder; lane 6: 2′-F-dATP+dTTPαS double modified PEAR amplified products; lane 7: control without PspGI; lane 8: control without KOD DNA polymerase; lane 9: control without 2′-F-dGTP and dTTPαS; lane 10: 20-bp DNA ladder.

    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.

    Techniques: Polyacrylamide Gel Electrophoresis, Electrophoresis, Amplification, Modification