phusion green hot start ii high fidelity dna polymerase kit  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Phusion Green Hot Start II High Fidelity DNA Polymerase 2 U µL
    Description:
    Thermo Scientific Phusion High Fidelity DNA Polymerases set a gold standard for high performance PCR Featuring an error rate 50 fold lower than that of Taq and 6 fold lower than that of Pfu Phusion High Fidelity DNA Polymerase is excellent choice for cloning and other applications requiring high fidelity Phusion DNA Polymerases offer robust performance with short protocol times even in the presence of PCR inhibitors and generate higher yields with lower enzyme amounts than other DNA polymerase With Phusion Hot Start II High Fidelity DNA Polymerase amplification proceeds without the production of nonspecific products due to the combination of Phusion DNA Polymerase and a reversibly bound specific Affibody ligand that inhibits DNA polymerase activity at room temperature The Affibody ligand also inhibits the 3 →5 exonuclease activity of the polymerase thus preventing degradation of primers and template DNA during reaction set up At temperatures that promote polymerase activity the ligand is released rendering the polymerase fully active Phusion Hot Start II DNA Polymerase is immediately reactivated at high temperatures so it does not require a separate activation step in PCR protocols Phusion Green Hot Start II High Fidelity DNA Polymerase is a combination of Phusion Hot Start II DNA Polymerase and the 5X Phusion Green Buffers The buffers include a density reagent and two tracking dyes which do not interfere with the robust performance of Phusion Hot Start II DNA Polymerase Researchers may directly load PCR products on a gel making this a versatile option for use in downstream applications such as DNA sequencing ligation and restriction digestion Highlights• Reaction set up at room temperature• No non specific amplification and primer degradation during reaction set up• Zero time reactivation due to unique hot start technology• High fidelity 52X Taq • Fast PCR due to short extension times 15 30 s kb • Robust reactions minimal optimization needed• Increased product yields with minimal enzyme amounts• Direct loading on gelsApplications• High fidelity PCR• High throughput• Amplification of difficult GC rich templates• Template generation for sequencing• Multiplex PCR• Long range PCR• Cloning• Mutagenesis• MicroarrayUsing Phusion DNA Polymerases Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases such as Taq DNA polymerases For optimal results use our Tm calculator at www thermofisher com tmcalculator
    Catalog Number:
    f537l
    Price:
    None
    Applications:
    High Fidelity PCR|Hot Start PCR|PCR|PCR & Real-Time PCR
    Category:
    Proteins Enzymes Peptides
    Buy from Supplier


    Structured Review

    Thermo Fisher phusion green hot start ii high fidelity dna polymerase kit
    Thermo Scientific Phusion High Fidelity DNA Polymerases set a gold standard for high performance PCR Featuring an error rate 50 fold lower than that of Taq and 6 fold lower than that of Pfu Phusion High Fidelity DNA Polymerase is excellent choice for cloning and other applications requiring high fidelity Phusion DNA Polymerases offer robust performance with short protocol times even in the presence of PCR inhibitors and generate higher yields with lower enzyme amounts than other DNA polymerase With Phusion Hot Start II High Fidelity DNA Polymerase amplification proceeds without the production of nonspecific products due to the combination of Phusion DNA Polymerase and a reversibly bound specific Affibody ligand that inhibits DNA polymerase activity at room temperature The Affibody ligand also inhibits the 3 →5 exonuclease activity of the polymerase thus preventing degradation of primers and template DNA during reaction set up At temperatures that promote polymerase activity the ligand is released rendering the polymerase fully active Phusion Hot Start II DNA Polymerase is immediately reactivated at high temperatures so it does not require a separate activation step in PCR protocols Phusion Green Hot Start II High Fidelity DNA Polymerase is a combination of Phusion Hot Start II DNA Polymerase and the 5X Phusion Green Buffers The buffers include a density reagent and two tracking dyes which do not interfere with the robust performance of Phusion Hot Start II DNA Polymerase Researchers may directly load PCR products on a gel making this a versatile option for use in downstream applications such as DNA sequencing ligation and restriction digestion Highlights• Reaction set up at room temperature• No non specific amplification and primer degradation during reaction set up• Zero time reactivation due to unique hot start technology• High fidelity 52X Taq • Fast PCR due to short extension times 15 30 s kb • Robust reactions minimal optimization needed• Increased product yields with minimal enzyme amounts• Direct loading on gelsApplications• High fidelity PCR• High throughput• Amplification of difficult GC rich templates• Template generation for sequencing• Multiplex PCR• Long range PCR• Cloning• Mutagenesis• MicroarrayUsing Phusion DNA Polymerases Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases such as Taq DNA polymerases For optimal results use our Tm calculator at www thermofisher com tmcalculator
    https://www.bioz.com/result/phusion green hot start ii high fidelity dna polymerase kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phusion green hot start ii high fidelity dna polymerase kit - by Bioz Stars, 2020-04
    99/100 stars

    Related Products / Commonly Used Together

    pcr

    Images

    Related Articles

    Clone Assay:

    Article Title: Role of tau in the spatial organization of axonal microtubules: keeping parallel microtubules evenly distributed despite macromolecular crowding
    Article Snippet: Paragraph title: Cloning of tau isoforms ... To obtain hTau24 three PCRs was necessary) using Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific).

    Article Title: Role of tau in the spatial organization of axonal microtubules: keeping parallel microtubules evenly distributed despite macromolecular crowding
    Article Snippet: .. Cloning of tau isoforms Tau 2N/4R and deltaNT cDNAs were obtained by amplifying hTau40 pET29b plasmid (catalogue no. 16316, Addgene, Cambridge, MA) either from the beginning or from P172 to L441 using Phusion Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific). .. The PCR products were first cloned into pENTR/D-TOPO by TOPO cloning (primers are listed in Supplemental Table 1) and then moved into pDEST17 by the LR reaction for recombinant protein expression with a His-Tag (Life Technologies).

    Article Title: Absolute measurement of gene transcripts with Selfie-digital PCR
    Article Snippet: Synthesis of DNA and RNA templates To test the accuracy of the Selfie-dPCR method, we cloned the coding DNA sequence of the GSK3β gene into a pJET 1.2 plasmid vector. .. The resulting cDNA was used as a template to amplify the Gsk3 β coding DNA sequence by using proofreading polymerase Phusion Green Hot Start II HF DNA Polymerase (F537S, Thermo-Fisher Scientific) and the following primers: forward PH-5′-ACCATGTCGGGGCGACCGAGAACC-3′ and reverse PH-5′-CCTGGGGGCTGTTCAGGTGGA-3′, where PH denotes phosphorylation at the 5′ end.

    Amplification:

    Article Title: HybriFree: a robust and rapid method for the development of monoclonal antibodies from different host species
    Article Snippet: .. PCR amplification of VH and VL regions PCR reactions were performed in 50 μl for a total of 35–40 cycles using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Life Technologies, US) with pre-optimized conditions for each reaction. .. The primers used for the amplification of antibody VH and VL regions were designed to maximally cover the variability of the VH and VL sequences.

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR. .. PCR amplified variable regions were gel purified (Qiagen, #28706) and combined with a CMV promoter containing DNA fragment, and the appropriate corresponding constant region DNA fragment (including a polyA tail) via overlapping PCR ( ).

    Article Title: Non-invasive continuous real-time in vivo analysis of microbial hydrogen production shows adaptation to fermentable carbohydrates in mice
    Article Snippet: The V4 region of 16S ribosomal RNA (rRNA) gene was amplified in duplicate PCR reactions for each sample in a total reaction volume of 50 μl. .. The master mix contained 1 μl of a unique barcoded primer, 515F-n and 806R-n (10 μM each per reaction), 1 μl dNTPs mixture, 0.5 μl Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U/μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl 5× Phusion Green HF Buffer, and 36.5 μl DNAse- RNAse-free water.

    Article Title: Age-Related Alterations Affecting the Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stromal Cells in an Equine Model
    Article Snippet: The PCR was performed by Phusion Green Hot Start II High-Fidelity DNA Polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA). .. Equine glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH ) was used as a housekeeping gene for amplification control during the PCR assay.

    Article Title: Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota, et al. Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota
    Article Snippet: In experiment 2, the V4 region within the 16S ribosomal RNA (rRNA) gene was amplified by primary PCR as triplicates using 505F and 806R primers (Caporaso et al., ). .. Primary PCR was carried out in a reaction mixture (reaction volume 50 μl) consisting of 1 μl each of 10 mmol/L dNTPs (Thermo scientific, MA, USA) 5 μl forward primer 505F (10 μmol/L; 5′–GTGCCAGCMGCCGCGGTAA‐3′) and 5 μl reverse primer 806R (10 μmol/L; 5′–GGACTACHVGGGTWTCTAAT‐3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High‐Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5× Green HF PCR buffer (F‐537), 5 μl template DNA and 23.5 μl sterile water.

    Article Title: Microbial Communities in Sediments From Four Mildly Acidic Ephemeral Salt Lakes in the Yilgarn Craton (Australia) – Terrestrial Analogs to Ancient Mars
    Article Snippet: Paragraph title: Illumina 16S Amplicon Sequencing ... PCR reactions were performed in triplicate using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Thermo Fisher Scientific, Sweden).

    Article Title: Urbanization Reduces Transfer of Diverse Environmental Microbiota Indoors
    Article Snippet: The V4 region within the 16S ribosomal RNA (rRNA) gene was amplified in triplicates during the primary PCRs using 515F and 806R primers (Caporaso et al., , ). .. The primary PCRs were carried out in 50 μl reaction volumes consisting of 1 μl each of 10 mM deoxynucleotide triphosphates (dNTPs; Thermo scientific, MA, USA), 5 μl forward primer 505F (10 μM; 5′ –GTGCCAGCMGCCGCGGTAA-3′) and 5 μl reverse primer 806R (10 μM; 5′-GGACTACHVGGGTWTCTAAT-3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High-Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5x Green HF PCR buffer (F-537), 5 μl of template DNA, and 23.5 μl sterile water.

    Positive Control:

    Article Title: Urbanization Reduces Transfer of Diverse Environmental Microbiota Indoors
    Article Snippet: The primary PCRs were carried out in 50 μl reaction volumes consisting of 1 μl each of 10 mM deoxynucleotide triphosphates (dNTPs; Thermo scientific, MA, USA), 5 μl forward primer 505F (10 μM; 5′ –GTGCCAGCMGCCGCGGTAA-3′) and 5 μl reverse primer 806R (10 μM; 5′-GGACTACHVGGGTWTCTAAT-3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High-Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5x Green HF PCR buffer (F-537), 5 μl of template DNA, and 23.5 μl sterile water. .. The PCR reactions were run in thermocycler (MJ Research, MA, USA) as follows: initial denaturation at 98°C for 5 min followed by 25 cycles of denaturation at 94°C for 1 min, annealing for 10 s at 50°C, extension for 1 min at 72°C, and then a final extension at 72°C for 10 min. A positive control (Cupriavidus necator JMP134, DSM 4058) was included in all PCRs, and sterile water was again used as a negative control to detect possible contaminations.

    Polymerase Chain Reaction:

    Article Title: HybriFree: a robust and rapid method for the development of monoclonal antibodies from different host species
    Article Snippet: .. PCR amplification of VH and VL regions PCR reactions were performed in 50 μl for a total of 35–40 cycles using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Life Technologies, US) with pre-optimized conditions for each reaction. .. The primers used for the amplification of antibody VH and VL regions were designed to maximally cover the variability of the VH and VL sequences.

    Article Title: Role of tau in the spatial organization of axonal microtubules: keeping parallel microtubules evenly distributed despite macromolecular crowding
    Article Snippet: The PCR products were first cloned into pENTR/D-TOPO by TOPO cloning (primers are listed in Supplemental Table 1) and then moved into pDEST17 by the LR reaction for recombinant protein expression with a His-Tag (Life Technologies). .. To obtain hTau24 three PCRs was necessary) using Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific).

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: Immunoglobulin heavy and light chain variable domain regions were PCR-amplified as previously described, using oligo-dT (ThermoScientific, #AM5730G) and SuperScriptIII (Invitrogen, #18080-044) ( ). .. Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR.

    Article Title: Non-invasive continuous real-time in vivo analysis of microbial hydrogen production shows adaptation to fermentable carbohydrates in mice
    Article Snippet: The V4 region of 16S ribosomal RNA (rRNA) gene was amplified in duplicate PCR reactions for each sample in a total reaction volume of 50 μl. .. The master mix contained 1 μl of a unique barcoded primer, 515F-n and 806R-n (10 μM each per reaction), 1 μl dNTPs mixture, 0.5 μl Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U/μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl 5× Phusion Green HF Buffer, and 36.5 μl DNAse- RNAse-free water.

    Article Title: Age-Related Alterations Affecting the Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stromal Cells in an Equine Model
    Article Snippet: .. The PCR was performed by Phusion Green Hot Start II High-Fidelity DNA Polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA). .. Equine glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH ) was used as a housekeeping gene for amplification control during the PCR assay.

    Article Title: A genome-wide map of hyper-edited RNA reveals numerous new sites
    Article Snippet: .. The PCR reaction was performed in C1000 Thermal Cycler (Bio-Rad) using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Thermo Scientific) and in a 20-μl reaction volume, according to the manufacturer’s instructions. ..

    Article Title: Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota, et al. Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota
    Article Snippet: .. Primary PCR was carried out in a reaction mixture (reaction volume 50 μl) consisting of 1 μl each of 10 mmol/L dNTPs (Thermo scientific, MA, USA) 5 μl forward primer 505F (10 μmol/L; 5′–GTGCCAGCMGCCGCGGTAA‐3′) and 5 μl reverse primer 806R (10 μmol/L; 5′–GGACTACHVGGGTWTCTAAT‐3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High‐Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5× Green HF PCR buffer (F‐537), 5 μl template DNA and 23.5 μl sterile water. .. The PCR reaction was performed in a thermocycler (MJ Research, MA, USA) as follows: initial denaturation at 98°C for 5 min, followed by 30 cycles (only 25 cycles was used for the moss sample) with denaturation at 94°C for 1 min, annealing for 10 s at 50°C and extension for 1 min at 72°C, and then a final extension at 72°C for 10 min. A positive (Cupriavidus necator JMP134, DSM 4058) and a negative controls (sterile water) were included in PCR and DNA was detected with agarose gel electrophoresis.

    Article Title: Microbial Communities in Sediments From Four Mildly Acidic Ephemeral Salt Lakes in the Yilgarn Craton (Australia) – Terrestrial Analogs to Ancient Mars
    Article Snippet: .. PCR reactions were performed in triplicate using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Thermo Fisher Scientific, Sweden). ..

    Article Title: Absolute measurement of gene transcripts with Selfie-digital PCR
    Article Snippet: The resulting cDNA was used as a template to amplify the Gsk3 β coding DNA sequence by using proofreading polymerase Phusion Green Hot Start II HF DNA Polymerase (F537S, Thermo-Fisher Scientific) and the following primers: forward PH-5′-ACCATGTCGGGGCGACCGAGAACC-3′ and reverse PH-5′-CCTGGGGGCTGTTCAGGTGGA-3′, where PH denotes phosphorylation at the 5′ end. .. The product was gel-purified (GeneJET Gel Extraction Kit, K0691, Thermo-Fisher Scientific) and cloned into the pJET1.2 vector (CloneJET PCR Cloning Kit, K1231, Thermo-Fisher Scientific).

    Electrophoresis:

    Article Title: Urbanization Reduces Transfer of Diverse Environmental Microbiota Indoors
    Article Snippet: The quality of the extracted DNA was checked using agarose gel (1.5%) electrophoresis and quantified with Quant-iT™ PicoGreen® dsDNA reagent kit (Thermo scientific, MA, USA). .. The primary PCRs were carried out in 50 μl reaction volumes consisting of 1 μl each of 10 mM deoxynucleotide triphosphates (dNTPs; Thermo scientific, MA, USA), 5 μl forward primer 505F (10 μM; 5′ –GTGCCAGCMGCCGCGGTAA-3′) and 5 μl reverse primer 806R (10 μM; 5′-GGACTACHVGGGTWTCTAAT-3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High-Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5x Green HF PCR buffer (F-537), 5 μl of template DNA, and 23.5 μl sterile water.

    Incubation:

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: After counting, cells were incubated with 8 μg of the autologous T/F gp120 per million cells and stained with: live/dead Aqua (Invitrogen, # L34957), anti-CD3 Pacific Blue (BD Pharmingen, # 558124), anti-CD19 BV650 (BioLegend, # 302237), anti-IgG FITC (BD Pharmingen, #555786), anti-His PE (Miltenyi Biotec, # 130-098-810), and single-cell sorted into 96-well plates containing 20 μl cell lysis buffer (Superscript III RT buffer, Tween, DTT, Invitrogen, #18080-044) (RNaseOUT, Invitrogen, #100000840). .. Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR.

    Expressing:

    Article Title: Role of tau in the spatial organization of axonal microtubules: keeping parallel microtubules evenly distributed despite macromolecular crowding
    Article Snippet: The PCR products were first cloned into pENTR/D-TOPO by TOPO cloning (primers are listed in Supplemental Table 1) and then moved into pDEST17 by the LR reaction for recombinant protein expression with a His-Tag (Life Technologies). .. To obtain hTau24 three PCRs was necessary) using Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific).

    Article Title: Age-Related Alterations Affecting the Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stromal Cells in an Equine Model
    Article Snippet: Paragraph title: 2.2. mRNA Expression of Mesenchymal and Self-Renewal Markers by RT-PCR and Real-Time PCR ... The PCR was performed by Phusion Green Hot Start II High-Fidelity DNA Polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA).

    High Performance Liquid Chromatography:

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: .. Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR. .. For the second round PCR reaction, 2.5 μl of first-round was used as a template.

    Sequencing:

    Article Title: Role of tau in the spatial organization of axonal microtubules: keeping parallel microtubules evenly distributed despite macromolecular crowding
    Article Snippet: To obtain hTau24 three PCRs was necessary) using Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific). .. Tau full length plasmid was used as template to design the primers and to amplify two fragments encoding hTau24 sequence (accession number NM_016834.4).

    Article Title: A genome-wide map of hyper-edited RNA reveals numerous new sites
    Article Snippet: Paragraph title: Direct sequencing validation of hyper-editing ... The PCR reaction was performed in C1000 Thermal Cycler (Bio-Rad) using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Thermo Scientific) and in a 20-μl reaction volume, according to the manufacturer’s instructions.

    Article Title: Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota, et al. Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota
    Article Snippet: Paragraph title: Sample preparation for MiSeq sequencing ... Primary PCR was carried out in a reaction mixture (reaction volume 50 μl) consisting of 1 μl each of 10 mmol/L dNTPs (Thermo scientific, MA, USA) 5 μl forward primer 505F (10 μmol/L; 5′–GTGCCAGCMGCCGCGGTAA‐3′) and 5 μl reverse primer 806R (10 μmol/L; 5′–GGACTACHVGGGTWTCTAAT‐3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High‐Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5× Green HF PCR buffer (F‐537), 5 μl template DNA and 23.5 μl sterile water.

    Article Title: Microbial Communities in Sediments From Four Mildly Acidic Ephemeral Salt Lakes in the Yilgarn Craton (Australia) – Terrestrial Analogs to Ancient Mars
    Article Snippet: Paragraph title: Illumina 16S Amplicon Sequencing ... PCR reactions were performed in triplicate using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Thermo Fisher Scientific, Sweden).

    Article Title: Urbanization Reduces Transfer of Diverse Environmental Microbiota Indoors
    Article Snippet: Paragraph title: Sample preparation for MiSeq sequencing ... The primary PCRs were carried out in 50 μl reaction volumes consisting of 1 μl each of 10 mM deoxynucleotide triphosphates (dNTPs; Thermo scientific, MA, USA), 5 μl forward primer 505F (10 μM; 5′ –GTGCCAGCMGCCGCGGTAA-3′) and 5 μl reverse primer 806R (10 μM; 5′-GGACTACHVGGGTWTCTAAT-3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High-Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5x Green HF PCR buffer (F-537), 5 μl of template DNA, and 23.5 μl sterile water.

    Article Title: Absolute measurement of gene transcripts with Selfie-digital PCR
    Article Snippet: .. The resulting cDNA was used as a template to amplify the Gsk3 β coding DNA sequence by using proofreading polymerase Phusion Green Hot Start II HF DNA Polymerase (F537S, Thermo-Fisher Scientific) and the following primers: forward PH-5′-ACCATGTCGGGGCGACCGAGAACC-3′ and reverse PH-5′-CCTGGGGGCTGTTCAGGTGGA-3′, where PH denotes phosphorylation at the 5′ end. .. The product was gel-purified (GeneJET Gel Extraction Kit, K0691, Thermo-Fisher Scientific) and cloned into the pJET1.2 vector (CloneJET PCR Cloning Kit, K1231, Thermo-Fisher Scientific).

    Recombinant:

    Article Title: Role of tau in the spatial organization of axonal microtubules: keeping parallel microtubules evenly distributed despite macromolecular crowding
    Article Snippet: The PCR products were first cloned into pENTR/D-TOPO by TOPO cloning (primers are listed in Supplemental Table 1) and then moved into pDEST17 by the LR reaction for recombinant protein expression with a His-Tag (Life Technologies). .. To obtain hTau24 three PCRs was necessary) using Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific).

    Staining:

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: After counting, cells were incubated with 8 μg of the autologous T/F gp120 per million cells and stained with: live/dead Aqua (Invitrogen, # L34957), anti-CD3 Pacific Blue (BD Pharmingen, # 558124), anti-CD19 BV650 (BioLegend, # 302237), anti-IgG FITC (BD Pharmingen, #555786), anti-His PE (Miltenyi Biotec, # 130-098-810), and single-cell sorted into 96-well plates containing 20 μl cell lysis buffer (Superscript III RT buffer, Tween, DTT, Invitrogen, #18080-044) (RNaseOUT, Invitrogen, #100000840). .. Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR.

    Article Title: Age-Related Alterations Affecting the Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stromal Cells in an Equine Model
    Article Snippet: The PCR was performed by Phusion Green Hot Start II High-Fidelity DNA Polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA). .. The amplified DNA was then electrophoresed on a 2% agarose gel and visualized by ethidium bromide staining; images were acquired by Image Station 2000R (Kodak, NY, USA).

    Purification:

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: .. Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR. .. For the second round PCR reaction, 2.5 μl of first-round was used as a template.

    Article Title: Non-invasive continuous real-time in vivo analysis of microbial hydrogen production shows adaptation to fermentable carbohydrates in mice
    Article Snippet: The master mix contained 1 μl of a unique barcoded primer, 515F-n and 806R-n (10 μM each per reaction), 1 μl dNTPs mixture, 0.5 μl Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U/μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl 5× Phusion Green HF Buffer, and 36.5 μl DNAse- RNAse-free water. .. The PCR product was visualised in 1% agarose gel (~290 bp) and purified with CleanPCR kit (CleanNA, Alphen aan den Rijn, The Netherlands).

    Article Title: Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota, et al. Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota
    Article Snippet: Primary PCR was carried out in a reaction mixture (reaction volume 50 μl) consisting of 1 μl each of 10 mmol/L dNTPs (Thermo scientific, MA, USA) 5 μl forward primer 505F (10 μmol/L; 5′–GTGCCAGCMGCCGCGGTAA‐3′) and 5 μl reverse primer 806R (10 μmol/L; 5′–GGACTACHVGGGTWTCTAAT‐3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High‐Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5× Green HF PCR buffer (F‐537), 5 μl template DNA and 23.5 μl sterile water. .. The PCR products were purified using Agencourt AMPure XP solution (Beckman Coulter Ins.).

    Article Title: Urbanization Reduces Transfer of Diverse Environmental Microbiota Indoors
    Article Snippet: The primary PCRs were carried out in 50 μl reaction volumes consisting of 1 μl each of 10 mM deoxynucleotide triphosphates (dNTPs; Thermo scientific, MA, USA), 5 μl forward primer 505F (10 μM; 5′ –GTGCCAGCMGCCGCGGTAA-3′) and 5 μl reverse primer 806R (10 μM; 5′-GGACTACHVGGGTWTCTAAT-3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High-Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5x Green HF PCR buffer (F-537), 5 μl of template DNA, and 23.5 μl sterile water. .. The PCR products were purified using Agencourt AMPure XP solution (Beckman Coulter Inc., 1:1 ratio of bead solution to PCR volume) and eluted with sterile water to minimize the carryover of primary PCR primers.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Age-Related Alterations Affecting the Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stromal Cells in an Equine Model
    Article Snippet: Paragraph title: 2.2. mRNA Expression of Mesenchymal and Self-Renewal Markers by RT-PCR and Real-Time PCR ... The PCR was performed by Phusion Green Hot Start II High-Fidelity DNA Polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA).

    Gel Extraction:

    Article Title: A genome-wide map of hyper-edited RNA reveals numerous new sites
    Article Snippet: The PCR reaction was performed in C1000 Thermal Cycler (Bio-Rad) using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Thermo Scientific) and in a 20-μl reaction volume, according to the manufacturer’s instructions. .. PCR products were run on a 1% agarose gel, extracted using MinElute gel extraction kit (Qiagen), and sequenced in Hy Laboratories.

    Article Title: Absolute measurement of gene transcripts with Selfie-digital PCR
    Article Snippet: The resulting cDNA was used as a template to amplify the Gsk3 β coding DNA sequence by using proofreading polymerase Phusion Green Hot Start II HF DNA Polymerase (F537S, Thermo-Fisher Scientific) and the following primers: forward PH-5′-ACCATGTCGGGGCGACCGAGAACC-3′ and reverse PH-5′-CCTGGGGGCTGTTCAGGTGGA-3′, where PH denotes phosphorylation at the 5′ end. .. The product was gel-purified (GeneJET Gel Extraction Kit, K0691, Thermo-Fisher Scientific) and cloned into the pJET1.2 vector (CloneJET PCR Cloning Kit, K1231, Thermo-Fisher Scientific).

    Nested PCR:

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: .. Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR. .. For the second round PCR reaction, 2.5 μl of first-round was used as a template.

    Agarose Gel Electrophoresis:

    Article Title: Non-invasive continuous real-time in vivo analysis of microbial hydrogen production shows adaptation to fermentable carbohydrates in mice
    Article Snippet: The master mix contained 1 μl of a unique barcoded primer, 515F-n and 806R-n (10 μM each per reaction), 1 μl dNTPs mixture, 0.5 μl Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U/μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl 5× Phusion Green HF Buffer, and 36.5 μl DNAse- RNAse-free water. .. The PCR product was visualised in 1% agarose gel (~290 bp) and purified with CleanPCR kit (CleanNA, Alphen aan den Rijn, The Netherlands).

    Article Title: Age-Related Alterations Affecting the Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stromal Cells in an Equine Model
    Article Snippet: The PCR was performed by Phusion Green Hot Start II High-Fidelity DNA Polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA). .. The amplified DNA was then electrophoresed on a 2% agarose gel and visualized by ethidium bromide staining; images were acquired by Image Station 2000R (Kodak, NY, USA).

    Article Title: A genome-wide map of hyper-edited RNA reveals numerous new sites
    Article Snippet: The PCR reaction was performed in C1000 Thermal Cycler (Bio-Rad) using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Thermo Scientific) and in a 20-μl reaction volume, according to the manufacturer’s instructions. .. PCR products were run on a 1% agarose gel, extracted using MinElute gel extraction kit (Qiagen), and sequenced in Hy Laboratories.

    Article Title: Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota, et al. Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota
    Article Snippet: Primary PCR was carried out in a reaction mixture (reaction volume 50 μl) consisting of 1 μl each of 10 mmol/L dNTPs (Thermo scientific, MA, USA) 5 μl forward primer 505F (10 μmol/L; 5′–GTGCCAGCMGCCGCGGTAA‐3′) and 5 μl reverse primer 806R (10 μmol/L; 5′–GGACTACHVGGGTWTCTAAT‐3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High‐Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5× Green HF PCR buffer (F‐537), 5 μl template DNA and 23.5 μl sterile water. .. The PCR reaction was performed in a thermocycler (MJ Research, MA, USA) as follows: initial denaturation at 98°C for 5 min, followed by 30 cycles (only 25 cycles was used for the moss sample) with denaturation at 94°C for 1 min, annealing for 10 s at 50°C and extension for 1 min at 72°C, and then a final extension at 72°C for 10 min. A positive (Cupriavidus necator JMP134, DSM 4058) and a negative controls (sterile water) were included in PCR and DNA was detected with agarose gel electrophoresis.

    Article Title: Urbanization Reduces Transfer of Diverse Environmental Microbiota Indoors
    Article Snippet: The quality of the extracted DNA was checked using agarose gel (1.5%) electrophoresis and quantified with Quant-iT™ PicoGreen® dsDNA reagent kit (Thermo scientific, MA, USA). .. The primary PCRs were carried out in 50 μl reaction volumes consisting of 1 μl each of 10 mM deoxynucleotide triphosphates (dNTPs; Thermo scientific, MA, USA), 5 μl forward primer 505F (10 μM; 5′ –GTGCCAGCMGCCGCGGTAA-3′) and 5 μl reverse primer 806R (10 μM; 5′-GGACTACHVGGGTWTCTAAT-3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High-Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5x Green HF PCR buffer (F-537), 5 μl of template DNA, and 23.5 μl sterile water.

    Plasmid Preparation:

    Article Title: Role of tau in the spatial organization of axonal microtubules: keeping parallel microtubules evenly distributed despite macromolecular crowding
    Article Snippet: Cloning of tau isoforms Tau 2N/4R and deltaNT cDNAs were obtained by amplifying hTau40 pET29b plasmid (catalogue no. 16316, Addgene, Cambridge, MA) either from the beginning or from P172 to L441 using Phusion Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific). .. To obtain hTau24 three PCRs was necessary) using Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific).

    Article Title: Role of tau in the spatial organization of axonal microtubules: keeping parallel microtubules evenly distributed despite macromolecular crowding
    Article Snippet: .. Cloning of tau isoforms Tau 2N/4R and deltaNT cDNAs were obtained by amplifying hTau40 pET29b plasmid (catalogue no. 16316, Addgene, Cambridge, MA) either from the beginning or from P172 to L441 using Phusion Hot Start II High-Fidelity DNA Polymerase (catalogue: F-537L, Thermo Fischer Scientific). .. The PCR products were first cloned into pENTR/D-TOPO by TOPO cloning (primers are listed in Supplemental Table 1) and then moved into pDEST17 by the LR reaction for recombinant protein expression with a His-Tag (Life Technologies).

    Article Title: Absolute measurement of gene transcripts with Selfie-digital PCR
    Article Snippet: Synthesis of DNA and RNA templates To test the accuracy of the Selfie-dPCR method, we cloned the coding DNA sequence of the GSK3β gene into a pJET 1.2 plasmid vector. .. The resulting cDNA was used as a template to amplify the Gsk3 β coding DNA sequence by using proofreading polymerase Phusion Green Hot Start II HF DNA Polymerase (F537S, Thermo-Fisher Scientific) and the following primers: forward PH-5′-ACCATGTCGGGGCGACCGAGAACC-3′ and reverse PH-5′-CCTGGGGGCTGTTCAGGTGGA-3′, where PH denotes phosphorylation at the 5′ end.

    Software:

    Article Title: Age-Related Alterations Affecting the Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stromal Cells in an Equine Model
    Article Snippet: The PCR was performed by Phusion Green Hot Start II High-Fidelity DNA Polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA). .. The primer sequences were designed by Primer design Software (Primer3-based OligoPerfect, Thermo Fisher Scientific, Waltham, MA, USA).

    Real-time Polymerase Chain Reaction:

    Article Title: Age-Related Alterations Affecting the Chondrogenic Differentiation of Synovial Fluid Mesenchymal Stromal Cells in an Equine Model
    Article Snippet: Paragraph title: 2.2. mRNA Expression of Mesenchymal and Self-Renewal Markers by RT-PCR and Real-Time PCR ... The PCR was performed by Phusion Green Hot Start II High-Fidelity DNA Polymerase kit (Thermo Fisher Scientific, Waltham, MA, USA).

    Binding Assay:

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: CD3+ gp120/His+ cells were monitored as positive controls for gp120 binding and detection. .. Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR.

    Sample Prep:

    Article Title: Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota, et al. Short‐term direct contact with soil and plant materials leads to an immediate increase in diversity of skin microbiota
    Article Snippet: Paragraph title: Sample preparation for MiSeq sequencing ... Primary PCR was carried out in a reaction mixture (reaction volume 50 μl) consisting of 1 μl each of 10 mmol/L dNTPs (Thermo scientific, MA, USA) 5 μl forward primer 505F (10 μmol/L; 5′–GTGCCAGCMGCCGCGGTAA‐3′) and 5 μl reverse primer 806R (10 μmol/L; 5′–GGACTACHVGGGTWTCTAAT‐3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High‐Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5× Green HF PCR buffer (F‐537), 5 μl template DNA and 23.5 μl sterile water.

    Article Title: Urbanization Reduces Transfer of Diverse Environmental Microbiota Indoors
    Article Snippet: Paragraph title: Sample preparation for MiSeq sequencing ... The primary PCRs were carried out in 50 μl reaction volumes consisting of 1 μl each of 10 mM deoxynucleotide triphosphates (dNTPs; Thermo scientific, MA, USA), 5 μl forward primer 505F (10 μM; 5′ –GTGCCAGCMGCCGCGGTAA-3′) and 5 μl reverse primer 806R (10 μM; 5′-GGACTACHVGGGTWTCTAAT-3′), 0.5 μl 2 U/μl Phusion Green Hot Start II High-Fidelity DNA polymerase (Thermo scientific, MA, USA), 10 μl 5x Green HF PCR buffer (F-537), 5 μl of template DNA, and 23.5 μl sterile water.

    Spectrophotometry:

    Article Title: Non-invasive continuous real-time in vivo analysis of microbial hydrogen production shows adaptation to fermentable carbohydrates in mice
    Article Snippet: DNA was eluted in 50 μl of DNAse- RNAse-free water and its concentration measured using a DS-11 FX+ Spectrophotometer/Fluorometer (DeNovix Inc., Wilmington, USA). .. The master mix contained 1 μl of a unique barcoded primer, 515F-n and 806R-n (10 μM each per reaction), 1 μl dNTPs mixture, 0.5 μl Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U/μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl 5× Phusion Green HF Buffer, and 36.5 μl DNAse- RNAse-free water.

    Concentration Assay:

    Article Title: Non-invasive continuous real-time in vivo analysis of microbial hydrogen production shows adaptation to fermentable carbohydrates in mice
    Article Snippet: DNA was eluted in 50 μl of DNAse- RNAse-free water and its concentration measured using a DS-11 FX+ Spectrophotometer/Fluorometer (DeNovix Inc., Wilmington, USA). .. The master mix contained 1 μl of a unique barcoded primer, 515F-n and 806R-n (10 μM each per reaction), 1 μl dNTPs mixture, 0.5 μl Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U/μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl 5× Phusion Green HF Buffer, and 36.5 μl DNAse- RNAse-free water.

    Article Title: Microbial Communities in Sediments From Four Mildly Acidic Ephemeral Salt Lakes in the Yilgarn Craton (Australia) – Terrestrial Analogs to Ancient Mars
    Article Snippet: If the concentration of the DNA extract was lower than 0.1 ng/μl it was processed undiluted. .. PCR reactions were performed in triplicate using Phusion Green Hot Start II High-Fidelity DNA Polymerase (Thermo Fisher Scientific, Sweden).

    DNA Purification:

    Article Title: Non-invasive continuous real-time in vivo analysis of microbial hydrogen production shows adaptation to fermentable carbohydrates in mice
    Article Snippet: DNA purification was performed with a customized kit (AS1220; Promega) using 250 μl of the final supernatant pool. .. The master mix contained 1 μl of a unique barcoded primer, 515F-n and 806R-n (10 μM each per reaction), 1 μl dNTPs mixture, 0.5 μl Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U/μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl 5× Phusion Green HF Buffer, and 36.5 μl DNAse- RNAse-free water.

    Lysis:

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: After counting, cells were incubated with 8 μg of the autologous T/F gp120 per million cells and stained with: live/dead Aqua (Invitrogen, # L34957), anti-CD3 Pacific Blue (BD Pharmingen, # 558124), anti-CD19 BV650 (BioLegend, # 302237), anti-IgG FITC (BD Pharmingen, #555786), anti-His PE (Miltenyi Biotec, # 130-098-810), and single-cell sorted into 96-well plates containing 20 μl cell lysis buffer (Superscript III RT buffer, Tween, DTT, Invitrogen, #18080-044) (RNaseOUT, Invitrogen, #100000840). .. Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR.

    FACS:

    Article Title: VH1-69 Utilizing Antibodies Are Capable of Mediating Non-neutralizing Fc-Mediated Effector Functions Against the Transmitted/Founder gp120
    Article Snippet: Paragraph title: B Cell Sorting ... Five microliters of the RT reaction was used to amplify heavy (IgG only), kappa, and lambda chain variable regions using high performance liquid chromatography (HPLC) purified primers as described in Liao et al. ( ) and Phusion Hotstart II High Fidelity DNA Polymerase (Thermo Scientific, #F537S) for first round nested PCR.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher phusion green hot start ii high fidelity dna polymerase kit
    Phusion Green Hot Start Ii High Fidelity Dna Polymerase Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phusion green hot start ii high fidelity dna polymerase kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phusion green hot start ii high fidelity dna polymerase kit - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    Image Search Results