phusion gc buffer  (Thermo Fisher)


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    Name:
    Phusion GC Buffer Pack
    Description:
    Two optimized buffers are available for Thermo Scientific Phusion High Fidelity DNA Polymerases • 5X Phusion HF Buffer F 518L F 538L Phusion Green F 520L Detergent free • 5X Phusion GC Buffer F 519L F 539L Phusion Green F 521L Detergent free The error rate of Phusion DNA Polymerase in HF Buffer 4 4 × 10 7 is lower than that in GC Buffer 9 5 × 10 7 Therefore the HF Buffer should be used as the default buffer for high fidelity amplification However GC Buffer can improve the performance of Phusion DNA Polymerase on some difficult or long templates such as GC rich templates or those with complex secondary structures For applications such as microarray or DHPLC where the DNA templates need to be free of detergents we recommend using detergent free reaction buffers 5X Phusion Green Buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel The colored buffers do not interfere with the enzyme performance and are compatible with downstream applications such as DNA sequencing ligation and restriction digestion Related ProductsPhusion GC Buffer Pack detergent freePhusion HF Buffer Pack detergent freePhusion HF Buffer PackPhusion Green HF Buffer PackPhusion Green GC Buffer Pack
    Catalog Number:
    f519l
    Price:
    None
    Applications:
    High Fidelity PCR|PCR|PCR & Real-Time PCR
    Category:
    Lab Reagents and Chemicals
    Buy from Supplier


    Structured Review

    Thermo Fisher phusion gc buffer
    Analysis of OE-PCR performance of three polymerases Taq, <t>Phusion</t> and Pfu using gel electrophoresis. Full-size 581-bp Fel d 4 gene is indicated by an arrow
    Two optimized buffers are available for Thermo Scientific Phusion High Fidelity DNA Polymerases • 5X Phusion HF Buffer F 518L F 538L Phusion Green F 520L Detergent free • 5X Phusion GC Buffer F 519L F 539L Phusion Green F 521L Detergent free The error rate of Phusion DNA Polymerase in HF Buffer 4 4 × 10 7 is lower than that in GC Buffer 9 5 × 10 7 Therefore the HF Buffer should be used as the default buffer for high fidelity amplification However GC Buffer can improve the performance of Phusion DNA Polymerase on some difficult or long templates such as GC rich templates or those with complex secondary structures For applications such as microarray or DHPLC where the DNA templates need to be free of detergents we recommend using detergent free reaction buffers 5X Phusion Green Buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel The colored buffers do not interfere with the enzyme performance and are compatible with downstream applications such as DNA sequencing ligation and restriction digestion Related ProductsPhusion GC Buffer Pack detergent freePhusion HF Buffer Pack detergent freePhusion HF Buffer PackPhusion Green HF Buffer PackPhusion Green GC Buffer Pack
    https://www.bioz.com/result/phusion gc buffer/product/Thermo Fisher
    Average 99 stars, based on 44 article reviews
    Price from $9.99 to $1999.99
    phusion gc buffer - by Bioz Stars, 2020-04
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    Images

    1) Product Images from "High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis"

    Article Title: High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis

    Journal: 3 Biotech

    doi: 10.1007/s13205-017-0745-2

    Analysis of OE-PCR performance of three polymerases Taq, Phusion and Pfu using gel electrophoresis. Full-size 581-bp Fel d 4 gene is indicated by an arrow
    Figure Legend Snippet: Analysis of OE-PCR performance of three polymerases Taq, Phusion and Pfu using gel electrophoresis. Full-size 581-bp Fel d 4 gene is indicated by an arrow

    Techniques Used: Overlap Extension Polymerase Chain Reaction, Nucleic Acid Electrophoresis

    Related Articles

    Transduction:

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: Chromosomal DNA was also extracted from HeLa cell clone N+2 and DMD myoblast clone 3C5, which were derived from cells stably transduced with dHV.68/5′3′.F50 DNA through random and AAVS1 -targeted DNA insertion, respectively. .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C.

    Clone Assay:

    Article Title: Reactivation of Chromosomally Integrated Human Herpesvirus-6 by Telomeric Circle Formation
    Article Snippet: Inverse PCR to detect short t-circles 100 ng of total genomic DNA was used to amplify short t-circles using a primer pair facing against each other (see ) and Phusion high-fidel ity master mix with GC buffer (Thermo scientific). .. Amplified PCR products were cloned into TOPO 2.1 vector and sequenced using M13 forward and reverse primers.

    Article Title: Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene ▿
    Article Snippet: Note that all of the subsequent PCRs were performed in 50-μl reaction mixtures containing 1× HF or GC buffer (Finnzymes protocol), 200 μM dNTPs, 0.5 μM each primer, 1 U of Phusion DNA polymerase (Finnzyme Oy), and 1 μl of template. .. Next, a 484-bp fragment of WL rdhA1 was cloned between the plasmid M13r sequence and the 16S rRNA gene fragment using the unique SacI and BamHI sites.

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: To this end, the vector pEGGsH6 was constructed by introducing a multiple cloning site, a tobacco etch virus (TEV) protease cleavage site, the open reading frame (ORF) of eGFP and a C-terminal hexahistidine (6xHis) tag in frame with the BamHI and the EcoRI sites of the vector pGEX1λT. .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo).

    Article Title: High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis
    Article Snippet: The reaction was carried out in a 25 µL volume containing 200 mM of each dNTP, 1 µL of oligomix (1.5 pmol per primer), 30 pmol of outer primers, and either 2.5 µL of 10× Phusion GC buffer with 1 U of Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific) or 10× Pfu buffer with 1 U of Pfu polymerase (Thermo Fisher Scientific) or 10 µL of Taq mix (CRIE, Russia). .. The products of Phusion and Pfu enzymes reactions were gel purified by Gel Extraction Centrifugal Filter Units (Merck Millipore, Darmstadt, Germany), followed by the dA tailing (Taq DNA polymerase, Evrogen, Russia) and cloning into pGEM® -T Easy vector (Promega, Madison, USA).

    Amplification:

    Article Title: Reactivation of Chromosomally Integrated Human Herpesvirus-6 by Telomeric Circle Formation
    Article Snippet: Inverse PCR to detect short t-circles 100 ng of total genomic DNA was used to amplify short t-circles using a primer pair facing against each other (see ) and Phusion high-fidel ity master mix with GC buffer (Thermo scientific). .. The following amplification cycles were used: Initial denaturation at 98°C for 2 minutes, 28 cycles of denaturation at 98°C for 30 seconds, primer annealing at 64°C for 30 seconds and primer extension at 72°C for 7 minutes.

    Article Title: Dicentric breakage at telomere fusions
    Article Snippet: Paragraph title: Amplification of the telomere–telomere fusions by PCR ... PCR reactions (30 μL) contained ∼10 ng of genomic DNA, 1× Phusion GC buffer, 3% DMSO, 200 μM each dNTP, 0.5 μM each primer, and 0.6 U of Phusion Polymerase (Finnzymes).

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo). .. PCR products were purified with a PCR purification kit (Avegene) and amplified with primers B_Bamoutf 5′-GTACTCCGAGACGGATCC-3′ and D_Xhooutr 5′-TACCCGCAGCGTGAGCTC-3′ using Phusion polymerase with GC buffer and 25 cycles of 15 s at 98°C, 30 s at 60°C and 45 s at 72°C.

    Article Title: The Role of Abcb5 Alleles in Susceptibility to Haloperidol-Induced Toxicity in Mice and Humans
    Article Snippet: The DNA was divided into three aliquots of equal quantity, and amplified separately using one of the following three DNA polymerases: Phusion HF (Finnzymes), Kapa HiFi (Kapa Biosystems), and Herculase (Agilent). .. For analysis of the SJL strain, Phusion GC (Finnzymes) was also used to amplify a separate aliquot.

    Article Title: High-throughput assessment of context-dependent effects of chromatin proteins
    Article Snippet: .. Establishment of TRIP plasmid library A 21-nt random barcode was added to the TRIP vector by PCR amplification with Phusion polymerase (2 U, Thermo Fisher) in GC buffer with 10 ng template plasmid, 500 nM forward and reverse primer (JvA168 and JvA169) and dNTPs (250 µM each) in a total volume of 100 µl. ..

    Article Title: Development of a Model System for Tick-Borne Flavivirus Persistence in HEK 293T Cells
    Article Snippet: Paragraph title: Oligonucleotides and PCR amplification of the LGTV genome from persistently infected cells. ... The PCR mixture contained 1× Phusion buffer (Thermo Scientific, Atlanta, GA), 1 U of Phusion DNA polymerase, the forward and reverse primers at 0.4 mM each, and 0.4 mM deoxynucleoside triphosphates.

    Article Title: Enterovirus D68 receptor requirements unveiled by haploid genetics
    Article Snippet: 4310900 947_RD2, ; 4310901348_RD2, ; 4310902042_RD2, ; 4310902284_RD2, ) using 22 primer sets (details available on request). cDNA was synthesized using the Thermoscript RT-PCR System (Invitrogen) followed by PCR using Phusion High-Fidelity PCR Master Mix with GC Buffer (Finnzymes). .. Amplification products were treated with ExoSAP-IT (Affimetrix), and sequencing reactions were performed using BigDye terminator reagent (Life Technologies) and analysis of product on ABI3700 automated sequencer.

    Article Title: Replication Errors Made During Oogenesis Lead to Detectable De Novo mtDNA Mutations in Zebrafish Oocytes with a Low mtDNA Copy Number
    Article Snippet: .. PCR amplification was performed using Phusion Hot Start II DNA polymerase in GC-buffer (ThermoScientific, Waltham, MA): 30 sec at 98°, followed by 40 cycles of 10 sec at 98° (denaturation), 20 sec at 58° (annealing), and 8 min at 72° (extension), with a final step for 10 min at 72°. .. The PCR product was checked using electrophoresis on a 1% agarose gel containing ethidium bromide, allowing also the detection of large deletions.

    Stable Transfection:

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: Stably expressed and correctly folded proteins gave rise to a green fluorescence when the randomly produced PCR products were combined with a green fluorescent protein (GFP) reporter ( ) (Supplementary Figure S1). .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo).

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: Chromosomal DNA was also extracted from HeLa cell clone N+2 and DMD myoblast clone 3C5, which were derived from cells stably transduced with dHV.68/5′3′.F50 DNA through random and AAVS1 -targeted DNA insertion, respectively. .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C.

    Synthesized:

    Article Title: Enterovirus D68 receptor requirements unveiled by haploid genetics
    Article Snippet: .. 4310900 947_RD2, ; 4310901348_RD2, ; 4310902042_RD2, ; 4310902284_RD2, ) using 22 primer sets (details available on request). cDNA was synthesized using the Thermoscript RT-PCR System (Invitrogen) followed by PCR using Phusion High-Fidelity PCR Master Mix with GC Buffer (Finnzymes). .. Amplification products were treated with ExoSAP-IT (Affimetrix), and sequencing reactions were performed using BigDye terminator reagent (Life Technologies) and analysis of product on ABI3700 automated sequencer.

    Construct:

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: To this end, the vector pEGGsH6 was constructed by introducing a multiple cloning site, a tobacco etch virus (TEV) protease cleavage site, the open reading frame (ORF) of eGFP and a C-terminal hexahistidine (6xHis) tag in frame with the BamHI and the EcoRI sites of the vector pGEX1λT. .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo).

    Real-time Polymerase Chain Reaction:

    Article Title: Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene ▿
    Article Snippet: Paragraph title: Development of a four-gene qPCR calibration standard. ... Note that all of the subsequent PCRs were performed in 50-μl reaction mixtures containing 1× HF or GC buffer (Finnzymes protocol), 200 μM dNTPs, 0.5 μM each primer, 1 U of Phusion DNA polymerase (Finnzyme Oy), and 1 μl of template.

    Incubation:

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C. .. When the PCR program reached the lower annealing temperature of 52°C, 28 additional cycles were performed by using the same cycling conditions except for the use of a constant annealing temperature of 52°C.

    Touchdown PCR:

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C. .. When the PCR program reached the lower annealing temperature of 52°C, 28 additional cycles were performed by using the same cycling conditions except for the use of a constant annealing temperature of 52°C.

    Modification:

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: Random polymerase chain reaction screen For the identification of soluble N-terminal fragments of human RecQL4, the random polymerase chain reaction (PCR) approach of Kawasaki and Inagaki ( ) was modified. .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo).

    Derivative Assay:

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: Chromosomal DNA was also extracted from HeLa cell clone N+2 and DMD myoblast clone 3C5, which were derived from cells stably transduced with dHV.68/5′3′.F50 DNA through random and AAVS1 -targeted DNA insertion, respectively. .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C.

    Article Title: Enterovirus D68 receptor requirements unveiled by haploid genetics
    Article Snippet: 4310900 947_RD2, ; 4310901348_RD2, ; 4310902042_RD2, ; 4310902284_RD2, ) using 22 primer sets (details available on request). cDNA was synthesized using the Thermoscript RT-PCR System (Invitrogen) followed by PCR using Phusion High-Fidelity PCR Master Mix with GC Buffer (Finnzymes). .. Derived trace files were trimmed from primer sequences and assembled using Bionumerics software version 6 (Applied- Maths).

    Hybridization:

    Article Title: Reactivation of Chromosomally Integrated Human Herpesvirus-6 by Telomeric Circle Formation
    Article Snippet: Inverse PCR to detect short t-circles 100 ng of total genomic DNA was used to amplify short t-circles using a primer pair facing against each other (see ) and Phusion high-fidel ity master mix with GC buffer (Thermo scientific). .. Amplified PCR products were run on 1% agarose gel and were used for Southern hybridization.

    Inverse PCR:

    Article Title: Reactivation of Chromosomally Integrated Human Herpesvirus-6 by Telomeric Circle Formation
    Article Snippet: .. Inverse PCR to detect short t-circles 100 ng of total genomic DNA was used to amplify short t-circles using a primer pair facing against each other (see ) and Phusion high-fidel ity master mix with GC buffer (Thermo scientific). .. The following amplification cycles were used: Initial denaturation at 98°C for 2 minutes, 28 cycles of denaturation at 98°C for 30 seconds, primer annealing at 64°C for 30 seconds and primer extension at 72°C for 7 minutes.

    Ligation:

    Article Title: High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis
    Article Snippet: The reaction was carried out in a 25 µL volume containing 200 mM of each dNTP, 1 µL of oligomix (1.5 pmol per primer), 30 pmol of outer primers, and either 2.5 µL of 10× Phusion GC buffer with 1 U of Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific) or 10× Pfu buffer with 1 U of Pfu polymerase (Thermo Fisher Scientific) or 10 µL of Taq mix (CRIE, Russia). .. Five positive clones resulting from the ligation of the product obtained by each polymerase were selected and sequenced by Applied Biosystems 3500 Series Genetic Analyzers (Thermo Fisher Scientific).

    Infection:

    Article Title: Development of a Model System for Tick-Borne Flavivirus Persistence in HEK 293T Cells
    Article Snippet: Paragraph title: Oligonucleotides and PCR amplification of the LGTV genome from persistently infected cells. ... The PCR mixture contained 1× Phusion buffer (Thermo Scientific, Atlanta, GA), 1 U of Phusion DNA polymerase, the forward and reverse primers at 0.4 mM each, and 0.4 mM deoxynucleoside triphosphates.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Identification of Novel NPRAP/?-Catenin-Interacting Proteins and the Direct Association of NPRAP with Dynamin 2
    Article Snippet: Paragraph title: RNA extraction and RT-PCR ... PCR reactions were performed with 2 µl of the RT product in a 50-µl mixture containing 0.4 mM dNTPs, 0.4 mM of each primer, 0.5% DMSO, Phusion GC buffer and 1 unit Phusion High-Fidelity DNA Polymerase (Finnzymes, Finland).

    Article Title: Enterovirus D68 receptor requirements unveiled by haploid genetics
    Article Snippet: .. 4310900 947_RD2, ; 4310901348_RD2, ; 4310902042_RD2, ; 4310902284_RD2, ) using 22 primer sets (details available on request). cDNA was synthesized using the Thermoscript RT-PCR System (Invitrogen) followed by PCR using Phusion High-Fidelity PCR Master Mix with GC Buffer (Finnzymes). .. Amplification products were treated with ExoSAP-IT (Affimetrix), and sequencing reactions were performed using BigDye terminator reagent (Life Technologies) and analysis of product on ABI3700 automated sequencer.

    DNA Sequencing:

    Article Title: The Role of Abcb5 Alleles in Susceptibility to Haloperidol-Induced Toxicity in Mice and Humans
    Article Snippet: After end repair of the DNA fragments and addition of “A” base to the 3′ ends, standard Illumina genomic DNA sequencing adaptors were ligated. .. For analysis of the SJL strain, Phusion GC (Finnzymes) was also used to amplify a separate aliquot.

    Sequencing:

    Article Title: Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene ▿
    Article Snippet: Note that all of the subsequent PCRs were performed in 50-μl reaction mixtures containing 1× HF or GC buffer (Finnzymes protocol), 200 μM dNTPs, 0.5 μM each primer, 1 U of Phusion DNA polymerase (Finnzyme Oy), and 1 μl of template. .. First, a 236-bp fragment of the WL rdhA2 gene was added simultaneously with a 352-bp fragment of the WL rdhA3 gene between the 16S rRNA fragment and the plasmid T7f sequence by using the unique NotI and XhoI restriction enzyme plasmid target sites.

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: Then, PCR reactions were performed on a template with the first 1280 bp of the ORF of RecQL4 (GeneBank NM_004260), using the sequence-specific forward primers RecQL4_159F (5′-GTACTCCGAGACGGATCCCAGGCCGGCGGCGGGCTC-3′) or RecQL4_178F (5′-GTACTCCGAGACGGATCCCGCAGCTCCGAGTCGCTCC-3′), corresponding to aa 54 and 60, respectively. .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo).

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: Next, samples containing 160 ng of genomic DNA were subjected to PCR with the AAVS1 -specific primer # 211 ( 5′-CAGGTCCACCCTCTGCTG-3′ ), together with primer # 651 ( 5′-TTCCTAACCCCAACACTTGC-3′ ) targeting the vector DNA (i.e. hEF1α promoter sequence). .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C.

    Article Title: Enterovirus D68 receptor requirements unveiled by haploid genetics
    Article Snippet: Paragraph title: Sequencing of Recent EV-D68 Strains Isolated from Clinical Specimen. ... 4310900 947_RD2, ; 4310901348_RD2, ; 4310902042_RD2, ; 4310902284_RD2, ) using 22 primer sets (details available on request). cDNA was synthesized using the Thermoscript RT-PCR System (Invitrogen) followed by PCR using Phusion High-Fidelity PCR Master Mix with GC Buffer (Finnzymes).

    Article Title: Replication Errors Made During Oogenesis Lead to Detectable De Novo mtDNA Mutations in Zebrafish Oocytes with a Low mtDNA Copy Number
    Article Snippet: Paragraph title: mtDNA amplification and sequencing ... PCR amplification was performed using Phusion Hot Start II DNA polymerase in GC-buffer (ThermoScientific, Waltham, MA): 30 sec at 98°, followed by 40 cycles of 10 sec at 98° (denaturation), 20 sec at 58° (annealing), and 8 min at 72° (extension), with a final step for 10 min at 72°.

    Sonication:

    Article Title: The Role of Abcb5 Alleles in Susceptibility to Haloperidol-Induced Toxicity in Mice and Humans
    Article Snippet: A total of 3 μg of DNA from each of nine inbred mouse strains was sonicated using a Covaris instrument with the following settings: duty cycle, 10%; intensity, 4; cycles per burst, 200; and time, 60 seconds. .. For analysis of the SJL strain, Phusion GC (Finnzymes) was also used to amplify a separate aliquot.

    Fluorescence:

    Article Title: Dicentric breakage at telomere fusions
    Article Snippet: PCR reactions (30 μL) contained ∼10 ng of genomic DNA, 1× Phusion GC buffer, 3% DMSO, 200 μM each dNTP, 0.5 μM each primer, and 0.6 U of Phusion Polymerase (Finnzymes). .. The products (10 μL) were separated through a 1% agarose gel containing 0.1 μg/mL ethidium bromide, and fluorescence was quantified using a Typhoon imager.

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: Stably expressed and correctly folded proteins gave rise to a green fluorescence when the randomly produced PCR products were combined with a green fluorescent protein (GFP) reporter ( ) (Supplementary Figure S1). .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo).

    Isolation:

    Article Title: Enterovirus D68 receptor requirements unveiled by haploid genetics
    Article Snippet: Paragraph title: Sequencing of Recent EV-D68 Strains Isolated from Clinical Specimen. ... 4310900 947_RD2, ; 4310901348_RD2, ; 4310902042_RD2, ; 4310902284_RD2, ) using 22 primer sets (details available on request). cDNA was synthesized using the Thermoscript RT-PCR System (Invitrogen) followed by PCR using Phusion High-Fidelity PCR Master Mix with GC Buffer (Finnzymes).

    Size-exclusion Chromatography:

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C. .. When the PCR program reached the lower annealing temperature of 52°C, 28 additional cycles were performed by using the same cycling conditions except for the use of a constant annealing temperature of 52°C.

    Article Title: Replication Errors Made During Oogenesis Lead to Detectable De Novo mtDNA Mutations in Zebrafish Oocytes with a Low mtDNA Copy Number
    Article Snippet: .. PCR amplification was performed using Phusion Hot Start II DNA polymerase in GC-buffer (ThermoScientific, Waltham, MA): 30 sec at 98°, followed by 40 cycles of 10 sec at 98° (denaturation), 20 sec at 58° (annealing), and 8 min at 72° (extension), with a final step for 10 min at 72°. .. The PCR product was checked using electrophoresis on a 1% agarose gel containing ethidium bromide, allowing also the detection of large deletions.

    Purification:

    Article Title: Identification of Novel NPRAP/?-Catenin-Interacting Proteins and the Direct Association of NPRAP with Dynamin 2
    Article Snippet: Purified RNA (1 µg) was reverse-transcribed with 200 units of SuperScript® II reverse transcriptase (Invitrogen, USA) for 1 hour (37°C) following the supplier's recommended protocol. .. PCR reactions were performed with 2 µl of the RT product in a 50-µl mixture containing 0.4 mM dNTPs, 0.4 mM of each primer, 0.5% DMSO, Phusion GC buffer and 1 unit Phusion High-Fidelity DNA Polymerase (Finnzymes, Finland).

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo). .. PCR products were purified with a PCR purification kit (Avegene) and amplified with primers B_Bamoutf 5′-GTACTCCGAGACGGATCC-3′ and D_Xhooutr 5′-TACCCGCAGCGTGAGCTC-3′ using Phusion polymerase with GC buffer and 25 cycles of 15 s at 98°C, 30 s at 60°C and 45 s at 72°C.

    Article Title: The Role of Abcb5 Alleles in Susceptibility to Haloperidol-Induced Toxicity in Mice and Humans
    Article Snippet: For analysis of the SJL strain, Phusion GC (Finnzymes) was also used to amplify a separate aliquot. .. For analysis of the SJL strain, Phusion GC (Finnzymes) was also used to amplify a separate aliquot.

    Article Title: High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis
    Article Snippet: The reaction was carried out in a 25 µL volume containing 200 mM of each dNTP, 1 µL of oligomix (1.5 pmol per primer), 30 pmol of outer primers, and either 2.5 µL of 10× Phusion GC buffer with 1 U of Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific) or 10× Pfu buffer with 1 U of Pfu polymerase (Thermo Fisher Scientific) or 10 µL of Taq mix (CRIE, Russia). .. The products of Phusion and Pfu enzymes reactions were gel purified by Gel Extraction Centrifugal Filter Units (Merck Millipore, Darmstadt, Germany), followed by the dA tailing (Taq DNA polymerase, Evrogen, Russia) and cloning into pGEM® -T Easy vector (Promega, Madison, USA).

    Article Title: Replication Errors Made During Oogenesis Lead to Detectable De Novo mtDNA Mutations in Zebrafish Oocytes with a Low mtDNA Copy Number
    Article Snippet: PCR amplification was performed using Phusion Hot Start II DNA polymerase in GC-buffer (ThermoScientific, Waltham, MA): 30 sec at 98°, followed by 40 cycles of 10 sec at 98° (denaturation), 20 sec at 58° (annealing), and 8 min at 72° (extension), with a final step for 10 min at 72°. .. Amplicons were purified using the Agencourt AMPure XP system (Beckman, Fullerton, CA), according to the manufacturer’s protocol.

    Polymerase Chain Reaction:

    Article Title: Identification of Novel NPRAP/?-Catenin-Interacting Proteins and the Direct Association of NPRAP with Dynamin 2
    Article Snippet: .. PCR reactions were performed with 2 µl of the RT product in a 50-µl mixture containing 0.4 mM dNTPs, 0.4 mM of each primer, 0.5% DMSO, Phusion GC buffer and 1 unit Phusion High-Fidelity DNA Polymerase (Finnzymes, Finland). .. The PCR conditions for both dynamins were 5 minutes (min) at 95°C, 30 cycles of 30 seconds (s) at 95°C, 30 s at 64°C, and 30 s at 72°C and 10 min at 72°C.

    Article Title: Reactivation of Chromosomally Integrated Human Herpesvirus-6 by Telomeric Circle Formation
    Article Snippet: Inverse PCR to detect short t-circles 100 ng of total genomic DNA was used to amplify short t-circles using a primer pair facing against each other (see ) and Phusion high-fidel ity master mix with GC buffer (Thermo scientific). .. Amplified PCR products were run on 1% agarose gel and were used for Southern hybridization.

    Article Title: Dicentric breakage at telomere fusions
    Article Snippet: .. PCR reactions (30 μL) contained ∼10 ng of genomic DNA, 1× Phusion GC buffer, 3% DMSO, 200 μM each dNTP, 0.5 μM each primer, and 0.6 U of Phusion Polymerase (Finnzymes). .. The products (10 μL) were separated through a 1% agarose gel containing 0.1 μg/mL ethidium bromide, and fluorescence was quantified using a Typhoon imager.

    Article Title: Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene ▿
    Article Snippet: Note that all of the subsequent PCRs were performed in 50-μl reaction mixtures containing 1× HF or GC buffer (Finnzymes protocol), 200 μM dNTPs, 0.5 μM each primer, 1 U of Phusion DNA polymerase (Finnzyme Oy), and 1 μl of template. .. The WL rdhA2 gene fragment had a 5′ NotI site and a 3′ NcoI site engineered by PCR with primers WLrdhA2NotIf and WLrdhA2NcoIr, while the WL rdhA3 fragment had a 5′ NcoI site and a 3′ XhoI site engineered with primers WLrdhA3NcoIf and WLrdhA3XhoIr (Table ).

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo). ..

    Article Title: High-throughput assessment of context-dependent effects of chromatin proteins
    Article Snippet: .. Establishment of TRIP plasmid library A 21-nt random barcode was added to the TRIP vector by PCR amplification with Phusion polymerase (2 U, Thermo Fisher) in GC buffer with 10 ng template plasmid, 500 nM forward and reverse primer (JvA168 and JvA169) and dNTPs (250 µM each) in a total volume of 100 µl. ..

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C. .. When the PCR program reached the lower annealing temperature of 52°C, 28 additional cycles were performed by using the same cycling conditions except for the use of a constant annealing temperature of 52°C.

    Article Title: High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis
    Article Snippet: DNA was assembled from fragments by one-step PCR. .. The reaction was carried out in a 25 µL volume containing 200 mM of each dNTP, 1 µL of oligomix (1.5 pmol per primer), 30 pmol of outer primers, and either 2.5 µL of 10× Phusion GC buffer with 1 U of Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific) or 10× Pfu buffer with 1 U of Pfu polymerase (Thermo Fisher Scientific) or 10 µL of Taq mix (CRIE, Russia).

    Article Title: Development of a Model System for Tick-Borne Flavivirus Persistence in HEK 293T Cells
    Article Snippet: .. The PCR mixture contained 1× Phusion buffer (Thermo Scientific, Atlanta, GA), 1 U of Phusion DNA polymerase, the forward and reverse primers at 0.4 mM each, and 0.4 mM deoxynucleoside triphosphates. .. PCR was performed for all fragments with a Bio-Rad Quad thermocycler under the following conditions: 95°C for 5 min; 30 cycles of denaturation at 95°C for 1 min, annealing at 61°C for 1 min, and extension 72°C for 3 min; and a final extension at 72°C for 7 min.

    Article Title: Enterovirus D68 receptor requirements unveiled by haploid genetics
    Article Snippet: .. 4310900 947_RD2, ; 4310901348_RD2, ; 4310902042_RD2, ; 4310902284_RD2, ) using 22 primer sets (details available on request). cDNA was synthesized using the Thermoscript RT-PCR System (Invitrogen) followed by PCR using Phusion High-Fidelity PCR Master Mix with GC Buffer (Finnzymes). .. Amplification products were treated with ExoSAP-IT (Affimetrix), and sequencing reactions were performed using BigDye terminator reagent (Life Technologies) and analysis of product on ABI3700 automated sequencer.

    Article Title: Replication Errors Made During Oogenesis Lead to Detectable De Novo mtDNA Mutations in Zebrafish Oocytes with a Low mtDNA Copy Number
    Article Snippet: .. PCR amplification was performed using Phusion Hot Start II DNA polymerase in GC-buffer (ThermoScientific, Waltham, MA): 30 sec at 98°, followed by 40 cycles of 10 sec at 98° (denaturation), 20 sec at 58° (annealing), and 8 min at 72° (extension), with a final step for 10 min at 72°. .. The PCR product was checked using electrophoresis on a 1% agarose gel containing ethidium bromide, allowing also the detection of large deletions.

    Nested PCR:

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C. .. Next, 1 µl of each PCR sample was subjected to a nested PCR with the AAVS1 - and vector DNA-specific primers #212 ( 5′-GCTTTGCCACCCTATGCTGAC-3′ ) and # 188 ( 5′-GGATCTGAGGAACCCCTAGTGATGG-3′ ), respectively.

    Plasmid Preparation:

    Article Title: Reactivation of Chromosomally Integrated Human Herpesvirus-6 by Telomeric Circle Formation
    Article Snippet: Inverse PCR to detect short t-circles 100 ng of total genomic DNA was used to amplify short t-circles using a primer pair facing against each other (see ) and Phusion high-fidel ity master mix with GC buffer (Thermo scientific). .. Amplified PCR products were cloned into TOPO 2.1 vector and sequenced using M13 forward and reverse primers.

    Article Title: Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene ▿
    Article Snippet: A pCR2.1 vector carrying the Dehalobacter 16S rRNA gene was used as the backbone for the new vector (Fig. ). .. Note that all of the subsequent PCRs were performed in 50-μl reaction mixtures containing 1× HF or GC buffer (Finnzymes protocol), 200 μM dNTPs, 0.5 μM each primer, 1 U of Phusion DNA polymerase (Finnzyme Oy), and 1 μl of template.

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: To this end, the vector pEGGsH6 was constructed by introducing a multiple cloning site, a tobacco etch virus (TEV) protease cleavage site, the open reading frame (ORF) of eGFP and a C-terminal hexahistidine (6xHis) tag in frame with the BamHI and the EcoRI sites of the vector pGEX1λT. .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo).

    Article Title: High-throughput assessment of context-dependent effects of chromatin proteins
    Article Snippet: .. Establishment of TRIP plasmid library A 21-nt random barcode was added to the TRIP vector by PCR amplification with Phusion polymerase (2 U, Thermo Fisher) in GC buffer with 10 ng template plasmid, 500 nM forward and reverse primer (JvA168 and JvA169) and dNTPs (250 µM each) in a total volume of 100 µl. ..

    Article Title: Targeted Chromosomal Insertion of Large DNA into the Human Genome by a Fiber-Modified High-Capacity Adenovirus-Based Vector System
    Article Snippet: Next, samples containing 160 ng of genomic DNA were subjected to PCR with the AAVS1 -specific primer # 211 ( 5′-CAGGTCCACCCTCTGCTG-3′ ), together with primer # 651 ( 5′-TTCCTAACCCCAACACTTGC-3′ ) targeting the vector DNA (i.e. hEF1α promoter sequence). .. PCR mixtures of 50 µl containing 200 µM dNTPs, 0.2 µM of each primer, 0.012 U/µl of Phusion High-Fidelity DNA polymerase (Finnzymes) and 1× GC buffer (Finnzymes) were placed in a DNA Engine Tetrad 2 thermal cycler (Bio-Rad) and a touchdown PCR program was initiated by a 3-min incubation at 98°C, followed by 19 cycles of 98°C for 20 sec, an annealing period of 30 sec with the temperature decreasing by 0.5°C every cycle from 62 to 52°C, and a 2-min extension step at 72°C.

    Article Title: High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis
    Article Snippet: The reaction was carried out in a 25 µL volume containing 200 mM of each dNTP, 1 µL of oligomix (1.5 pmol per primer), 30 pmol of outer primers, and either 2.5 µL of 10× Phusion GC buffer with 1 U of Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific) or 10× Pfu buffer with 1 U of Pfu polymerase (Thermo Fisher Scientific) or 10 µL of Taq mix (CRIE, Russia). .. The products of Phusion and Pfu enzymes reactions were gel purified by Gel Extraction Centrifugal Filter Units (Merck Millipore, Darmstadt, Germany), followed by the dA tailing (Taq DNA polymerase, Evrogen, Russia) and cloning into pGEM® -T Easy vector (Promega, Madison, USA).

    Software:

    Article Title: Enterovirus D68 receptor requirements unveiled by haploid genetics
    Article Snippet: 4310900 947_RD2, ; 4310901348_RD2, ; 4310902042_RD2, ; 4310902284_RD2, ) using 22 primer sets (details available on request). cDNA was synthesized using the Thermoscript RT-PCR System (Invitrogen) followed by PCR using Phusion High-Fidelity PCR Master Mix with GC Buffer (Finnzymes). .. Derived trace files were trimmed from primer sequences and assembled using Bionumerics software version 6 (Applied- Maths).

    Electrophoresis:

    Article Title: Replication Errors Made During Oogenesis Lead to Detectable De Novo mtDNA Mutations in Zebrafish Oocytes with a Low mtDNA Copy Number
    Article Snippet: PCR amplification was performed using Phusion Hot Start II DNA polymerase in GC-buffer (ThermoScientific, Waltham, MA): 30 sec at 98°, followed by 40 cycles of 10 sec at 98° (denaturation), 20 sec at 58° (annealing), and 8 min at 72° (extension), with a final step for 10 min at 72°. .. The PCR product was checked using electrophoresis on a 1% agarose gel containing ethidium bromide, allowing also the detection of large deletions.

    RNA Extraction:

    Article Title: Identification of Novel NPRAP/?-Catenin-Interacting Proteins and the Direct Association of NPRAP with Dynamin 2
    Article Snippet: Paragraph title: RNA extraction and RT-PCR ... PCR reactions were performed with 2 µl of the RT product in a 50-µl mixture containing 0.4 mM dNTPs, 0.4 mM of each primer, 0.5% DMSO, Phusion GC buffer and 1 unit Phusion High-Fidelity DNA Polymerase (Finnzymes, Finland).

    Agarose Gel Electrophoresis:

    Article Title: Reactivation of Chromosomally Integrated Human Herpesvirus-6 by Telomeric Circle Formation
    Article Snippet: Inverse PCR to detect short t-circles 100 ng of total genomic DNA was used to amplify short t-circles using a primer pair facing against each other (see ) and Phusion high-fidel ity master mix with GC buffer (Thermo scientific). .. Amplified PCR products were run on 1% agarose gel and were used for Southern hybridization.

    Article Title: Dicentric breakage at telomere fusions
    Article Snippet: PCR reactions (30 μL) contained ∼10 ng of genomic DNA, 1× Phusion GC buffer, 3% DMSO, 200 μM each dNTP, 0.5 μM each primer, and 0.6 U of Phusion Polymerase (Finnzymes). .. The products (10 μL) were separated through a 1% agarose gel containing 0.1 μg/mL ethidium bromide, and fluorescence was quantified using a Typhoon imager.

    Article Title: The Role of Abcb5 Alleles in Susceptibility to Haloperidol-Induced Toxicity in Mice and Humans
    Article Snippet: For analysis of the SJL strain, Phusion GC (Finnzymes) was also used to amplify a separate aliquot. .. For analysis of the SJL strain, Phusion GC (Finnzymes) was also used to amplify a separate aliquot.

    Article Title: Replication Errors Made During Oogenesis Lead to Detectable De Novo mtDNA Mutations in Zebrafish Oocytes with a Low mtDNA Copy Number
    Article Snippet: PCR amplification was performed using Phusion Hot Start II DNA polymerase in GC-buffer (ThermoScientific, Waltham, MA): 30 sec at 98°, followed by 40 cycles of 10 sec at 98° (denaturation), 20 sec at 58° (annealing), and 8 min at 72° (extension), with a final step for 10 min at 72°. .. The PCR product was checked using electrophoresis on a 1% agarose gel containing ethidium bromide, allowing also the detection of large deletions.

    Next-Generation Sequencing:

    Article Title: The Role of Abcb5 Alleles in Susceptibility to Haloperidol-Induced Toxicity in Mice and Humans
    Article Snippet: Paragraph title: Next-Generation Sequencing of the Genome of Nine Inbred Mouse Strains ... For analysis of the SJL strain, Phusion GC (Finnzymes) was also used to amplify a separate aliquot.

    Produced:

    Article Title: The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding
    Article Snippet: Stably expressed and correctly folded proteins gave rise to a green fluorescence when the randomly produced PCR products were combined with a green fluorescent protein (GFP) reporter ( ) (Supplementary Figure S1). .. PCR was performed in 50 μl reactions with Phusion DNA polymerase and GC buffer (Finnzymes/Thermo).

    Concentration Assay:

    Article Title: High-throughput assessment of context-dependent effects of chromatin proteins
    Article Snippet: Establishment of TRIP plasmid library A 21-nt random barcode was added to the TRIP vector by PCR amplification with Phusion polymerase (2 U, Thermo Fisher) in GC buffer with 10 ng template plasmid, 500 nM forward and reverse primer (JvA168 and JvA169) and dNTPs (250 µM each) in a total volume of 100 µl. .. The reaction was stopped by adding EDTA to a final concentration of 10 mM and heat inactivation for 20 min at 75 °C.

    Gel Extraction:

    Article Title: High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis
    Article Snippet: The reaction was carried out in a 25 µL volume containing 200 mM of each dNTP, 1 µL of oligomix (1.5 pmol per primer), 30 pmol of outer primers, and either 2.5 µL of 10× Phusion GC buffer with 1 U of Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific) or 10× Pfu buffer with 1 U of Pfu polymerase (Thermo Fisher Scientific) or 10 µL of Taq mix (CRIE, Russia). .. The products of Phusion and Pfu enzymes reactions were gel purified by Gel Extraction Centrifugal Filter Units (Merck Millipore, Darmstadt, Germany), followed by the dA tailing (Taq DNA polymerase, Evrogen, Russia) and cloning into pGEM® -T Easy vector (Promega, Madison, USA).

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    Thermo Fisher phusion gc buffer
    Analysis of OE-PCR performance of three polymerases Taq, <t>Phusion</t> and Pfu using gel electrophoresis. Full-size 581-bp Fel d 4 gene is indicated by an arrow
    Phusion Gc Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 508 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phusion gc buffer/product/Thermo Fisher
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    Thermo Fisher 1x phusion gc buffer
    Analysis of OE-PCR performance of three polymerases Taq, <t>Phusion</t> and Pfu using gel electrophoresis. Full-size 581-bp Fel d 4 gene is indicated by an arrow
    1x Phusion Gc Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1x phusion gc buffer/product/Thermo Fisher
    Average 93 stars, based on 8 article reviews
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    Analysis of OE-PCR performance of three polymerases Taq, Phusion and Pfu using gel electrophoresis. Full-size 581-bp Fel d 4 gene is indicated by an arrow

    Journal: 3 Biotech

    Article Title: High-fidelity PCR enzyme with DNA-binding domain facilitates de novo gene synthesis

    doi: 10.1007/s13205-017-0745-2

    Figure Lengend Snippet: Analysis of OE-PCR performance of three polymerases Taq, Phusion and Pfu using gel electrophoresis. Full-size 581-bp Fel d 4 gene is indicated by an arrow

    Article Snippet: The reaction was carried out in a 25 µL volume containing 200 mM of each dNTP, 1 µL of oligomix (1.5 pmol per primer), 30 pmol of outer primers, and either 2.5 µL of 10× Phusion GC buffer with 1 U of Phusion High-Fidelity DNA polymerase (Thermo Fisher Scientific) or 10× Pfu buffer with 1 U of Pfu polymerase (Thermo Fisher Scientific) or 10 µL of Taq mix (CRIE, Russia).

    Techniques: Overlap Extension Polymerase Chain Reaction, Nucleic Acid Electrophoresis