long range phusion pcr  (Thermo Fisher)


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    Name:
    Phusion Flash High Fidelity PCR Master Mix
    Description:
    Thermo Scientific Phusion Flash PCR Master Mix was developed to save valuable laboratory time Ready to use 2X master mix preserves the fidelity and the yield in the reaction when using extremely short PCR protocols Additionally the user only needs to add template and primers minimizing the number of pipetting steps The unique master mix composition permits usage of extremely short cycling protocols with both low and high complexity DNA templates 15 seconds per kilobase or less The master mix utilizes Phusion Flash II DNA Polymerase a modified proofreading DNA polymerase derived from Phusion Hot Start II High Fidelity DNA Polymerase Features• Extreme speed extension times of 15 s kb or less• Hot start modification allowing “zero time reactivation • Accuracy proofreading DNA polymerase with a fidelity of 25X Taq• High yields in reduced timeApplications• Fast PCR• High fidelity PCR• 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:
    f548l
    Price:
    None
    Category:
    Proteins Enzymes Peptides
    Applications:
    High Fidelity PCR|PCR|PCR & Real-Time PCR
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    Structured Review

    Thermo Fisher long range phusion pcr
    Thermo Scientific Phusion Flash PCR Master Mix was developed to save valuable laboratory time Ready to use 2X master mix preserves the fidelity and the yield in the reaction when using extremely short PCR protocols Additionally the user only needs to add template and primers minimizing the number of pipetting steps The unique master mix composition permits usage of extremely short cycling protocols with both low and high complexity DNA templates 15 seconds per kilobase or less The master mix utilizes Phusion Flash II DNA Polymerase a modified proofreading DNA polymerase derived from Phusion Hot Start II High Fidelity DNA Polymerase Features• Extreme speed extension times of 15 s kb or less• Hot start modification allowing “zero time reactivation • Accuracy proofreading DNA polymerase with a fidelity of 25X Taq• High yields in reduced timeApplications• Fast PCR• High fidelity PCR• 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/long range phusion pcr/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    long range phusion pcr - by Bioz Stars, 2021-03
    99/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: Altered intestinal microbiota–host mitochondria crosstalk in new onset Crohn's disease
    Article Snippet: The first PCR added the Illumina paired-end sequencing adaptors and the barcode sequences using modified universal 16S rDNA-V6 primers ( , ). .. Each reaction was prepared in a total volume of 50 μl using 50 ng of the extracted DNA, 0.5 μM of each primer and 1 × Phusion Flash High-Fidelity PCR Master Mix (Thermo Scientific, Vilnius, Lithuania). .. The second PCR was carried out using 10 μl of the first PCR products in a final volume of 50 μl using the primers PCRFWD1/PCRRVS1 ( ).

    Article Title: Novel reporter systems for facile evaluation of I-SceI-mediated genome editing
    Article Snippet: EGFP+ D-17 cells generated by gene correction were purified through FACS sorting to > 90% EGFP+. .. Genomic DNA was extracted (Gentra Puregene Cell Kit, QIAGEN) and the gene target was amplified by PCR using the Phusion™ Flash High-Fidelity PCR Master Mix (Finnzymes) with the following primers: hPGK-forward 5′-CAC GTC GGC AGT CGG CTC CCT CG-3′ and EGFP target-reverse 5′-CTC GTC CAT GCC GAG AGT GAT C-3′. .. The amplicon (799 bp) was subcloned into a pCR4-TOPO® vector (Invitrogen) following the manufacturer's instructions and then sequenced using standard procedures.

    Article Title: Time-Resolved Tracking of Mutations Reveals Diverse Allele Dynamics during Escherichia coli Antimicrobial Adaptive Evolution to Single Drugs and Drug Pairs
    Article Snippet: Amplicon Library Preparation and Sequencing All samples were amplified using the primers listed in Supplementary Table according to the descriptions in the above section for each drug condition. .. Each PCR reaction was performed in 0.2-mL sterile PCR tubes that contained 10-μL Phusion Flash High-Fidelity PCR Master Mix (Thermo Fisher Scientific, United States), 0.5 μM forward primer, 0.5 μM reverse primer, 1 μL DNA template and H2 0 to a total reaction volume of 20 μL. .. The PCR amplification consisted of an initial denaturation at 98°C for 30 s, 30 cycles consisting of 98°C for 10 s denaturation, 65°C for 10 s annealing, 72°C for 15 s elongation, 72°C for 60 s for final extension, and subsequent holding at 4°C.

    Article Title: Structure of the competence pilus major pilin ComGC in Streptococcus pneumoniae
    Article Snippet: In vitro ComGC processing The plasmids pJWV25-PilD and pACYCDuet-1-flcomGC were constructed as follows. .. Full-length pilD and full-length comGC were amplified from S. pneumoniae TIGR4 genomic DNA using Phusion Flash High-Fidelity PCR Master Mix (Thermo Fisher Scientific) and suitable primers ( supplemental Table S2 ). .. PCR products were digested with NotI (pilD ) or NdeI and Xho (comGC ) and subcloned into pJWV25 and pACYCDuet-1, respectively.

    Article Title: Immortalization of primary microglia: a new platform to study HIV regulation in the central nervous system
    Article Snippet: Similarly, CYCT1 from macaque microglial cells was amplified for purified DNA and macaque CYCT1-specific primers Fwd 5′-ACA GGG AAA CAG TCC ACC AG-3′ and Rvs 5′-TAT GAT TTA TCT GAT AGT-3′. .. In each case, we used the Phusion Flash High-Fidelity (ThermoFisher F548L) polymerase, and the following PCR program: initial denaturing at 98 °C for 10 s, 30 cycles of denaturing at 98 °C for 1 s, annealing at 62 °C for 5 s, and extension at 72 °C for 15 s/Kb of product, and a final extension step at 72 °C for 1 min. .. Expression of microglial cell surface markers Surface expression of microglia specific markers was detected by fluorescence microscopy.

    Article Title: Horizontal Transfer of DNA from the Mitochondrial to the Plastid Genome and Its Subsequent Evolution in Milkweeds (Apocynaceae)
    Article Snippet: We aligned assembly contigs and consensus sequences with a selection of asterid plastome sequences available in GenBank ( ) using MAFFT v. 6.857 ( ) and compared them in BioEdit v. 7.0.5.3 ( ) to determine insert sizes (length of nonhomologous regions). .. We confirmed the 5′ and 3′ ends of the insert sequence in the A. syriaca plastome by PCR (1× Phusion® Flash High-Fidelity PCR Master Mix [Finnzymes], 0.2 µM forward primer 5′-ACACTCTCGTAGCGCCGTATAGTCTT-3′, 0.2 µM reverse primer 5′-GGTTCAAAGGATTAGTGCACCCTTCA-3′, cycling conditions: 30 s 98 °C, 25 cycles of 10 s 98 °C, 30 s 61 °C, 90 s 72 °C, extension 10 min 72 °C) followed by standard Sanger dye-termination sequencing. .. We assembled the A. nivea plastome sequence using a combination of de novo and reference-guided assembly following ( ) and using the A. syriaca reference (GenBank: JF433943.1).

    Purification:

    Article Title: Phenotypic characterization of Gardnerella vaginalis subgroups suggests differences in their virulence potential
    Article Snippet: Sialidase activity was normalized to the OD of bacterial cultures. .. Purified genomic DNA from G . vaginalis isolates 47.3, 58.4, 60.1, 79.2, 86.1, and 114.2 was used to amplify the full-length sld gene with primers Sia5-F and Sia3-R with Phusion Flash high-fidelity master mix (Thermo Fisher Scientific). ..

    Concentration Assay:

    Article Title: A novel proof of concept for capturing the diversity of endophytic fungi preserved in herbarium specimens
    Article Snippet: In PCR1, we used phase-shifted primers ITS1F and ITS4 with universal sequences CS1 and CS2 attached (Integrated DNA Technologies Inc., USA). .. Each 20 µl reaction contained 10 µl of Phusion Flash High Fidelity Master Mix (Thermo Scientific, USA), 0.2 µl of 0.5 µM of each primer, 1 µl of BSA at a concentration of 20 mg ml−1 , 5 µl of DNA template and 3.6 µl of molecular biology grade water (Fisher Scientific, USA). .. The reaction mixture was amplified by PCR as follows: 98°C for 10 s, 28 cycles of 98°C for 1 s, 57°C for 5 s, 72°C for 20 s, and a 1 min extension at 72°C [ ].

    Amplification:

    Article Title: Novel reporter systems for facile evaluation of I-SceI-mediated genome editing
    Article Snippet: EGFP+ D-17 cells generated by gene correction were purified through FACS sorting to > 90% EGFP+. .. Genomic DNA was extracted (Gentra Puregene Cell Kit, QIAGEN) and the gene target was amplified by PCR using the Phusion™ Flash High-Fidelity PCR Master Mix (Finnzymes) with the following primers: hPGK-forward 5′-CAC GTC GGC AGT CGG CTC CCT CG-3′ and EGFP target-reverse 5′-CTC GTC CAT GCC GAG AGT GAT C-3′. .. The amplicon (799 bp) was subcloned into a pCR4-TOPO® vector (Invitrogen) following the manufacturer's instructions and then sequenced using standard procedures.

    Article Title: Structure of the competence pilus major pilin ComGC in Streptococcus pneumoniae
    Article Snippet: In vitro ComGC processing The plasmids pJWV25-PilD and pACYCDuet-1-flcomGC were constructed as follows. .. Full-length pilD and full-length comGC were amplified from S. pneumoniae TIGR4 genomic DNA using Phusion Flash High-Fidelity PCR Master Mix (Thermo Fisher Scientific) and suitable primers ( supplemental Table S2 ). .. PCR products were digested with NotI (pilD ) or NdeI and Xho (comGC ) and subcloned into pJWV25 and pACYCDuet-1, respectively.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Novel reporter systems for facile evaluation of I-SceI-mediated genome editing
    Article Snippet: EGFP+ D-17 cells generated by gene correction were purified through FACS sorting to > 90% EGFP+. .. Genomic DNA was extracted (Gentra Puregene Cell Kit, QIAGEN) and the gene target was amplified by PCR using the Phusion™ Flash High-Fidelity PCR Master Mix (Finnzymes) with the following primers: hPGK-forward 5′-CAC GTC GGC AGT CGG CTC CCT CG-3′ and EGFP target-reverse 5′-CTC GTC CAT GCC GAG AGT GAT C-3′. .. The amplicon (799 bp) was subcloned into a pCR4-TOPO® vector (Invitrogen) following the manufacturer's instructions and then sequenced using standard procedures.

    Sequencing:

    Article Title: Horizontal Transfer of DNA from the Mitochondrial to the Plastid Genome and Its Subsequent Evolution in Milkweeds (Apocynaceae)
    Article Snippet: We aligned assembly contigs and consensus sequences with a selection of asterid plastome sequences available in GenBank ( ) using MAFFT v. 6.857 ( ) and compared them in BioEdit v. 7.0.5.3 ( ) to determine insert sizes (length of nonhomologous regions). .. We confirmed the 5′ and 3′ ends of the insert sequence in the A. syriaca plastome by PCR (1× Phusion® Flash High-Fidelity PCR Master Mix [Finnzymes], 0.2 µM forward primer 5′-ACACTCTCGTAGCGCCGTATAGTCTT-3′, 0.2 µM reverse primer 5′-GGTTCAAAGGATTAGTGCACCCTTCA-3′, cycling conditions: 30 s 98 °C, 25 cycles of 10 s 98 °C, 30 s 61 °C, 90 s 72 °C, extension 10 min 72 °C) followed by standard Sanger dye-termination sequencing. .. We assembled the A. nivea plastome sequence using a combination of de novo and reference-guided assembly following ( ) and using the A. syriaca reference (GenBank: JF433943.1).

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  • 99
    Thermo Fisher long range phusion pcr
    Long Range Phusion Pcr, 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
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