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    Wednesday, 29 May 2013 16:18

    AccuPower ProFi Taq PCR PreMix

    AccuPower® ProFi Taq PCR PreMix for high efficiency and amplification of long range PCR.

    ProFiTaqPCRPreMix f01

    AccuPower® ProFi Taq PCR PreMix is a convenient lyophilized PCR master mix containing ProFi Taq DNA polymerase, reaction buffer, dNTPs, tracking dye, and a patented stabilizer. ProFi Taq DNA polymerase in the premix is a unique recombinant Taq DNA polymerase that offers enhanced amplification efficiency and higher fidelity for PCR. AccuPower® ProFi Taq PCR PreMix is applicable to any template DNA, and especially effective in amplifying large genomic DNA fragments around 20 kb. AccuPower® ProFi Taq PCR PreMix provides accurate long-range amplification of standard and amplification of low-copy target, and is highly suitable for all PCR applications.

     

    Features and Benefits

    Long PCR: ProFi Taq is especially effective in amplifying large genomic DNA fragments around 20 kb and amplifying Lambda DNA up to 30kb.
    Easy to use: All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized "PreMix" form.
    Reproducibility: Gentaur's strict quality controlled production system ensures that your results will be reproducible experiment after experiment.
    Convenient: Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided
    Stability: Stable at room temperature for a month and for 2 years in a -20°C freezer

     

    Specifications

    5' to 3' exonuclease: Yes
    3' to 5' exonuclease: Yes
    3' – A Overhang: Yes
    PCR product size: ~ 30kb


    Application

    - Primer extension
    - long-range amplification from genomic DNA
    - High amplification efficiency
    - Excellent performance on difficult templates
    - Amplification of low-copy targets
    - High yield and high sensitivity PCR

    Experimental data

    ProFi Taq fig1

    Figure 1. Comparison of PCR amplification efficiency between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5min, 30 cycles of 95°C for 20 sec, 55°C for 20 sec and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
    Target : human insulin receptor gene.
    Lane M : 100 bp DNA Ladder(Gentaur, Cat. No. D-1030)
    Lane 1 : 10 ng of human genomic DNA
    Lane 2 : 1 ng of human genomic DNA
    Lane 3 : 100 pg of human genomic DNA
    Lane 4 : 10 pg of human genomic DNA

    ProFi Taq fig2
    Figure 2. Comparison of PCR amplification efficiency between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    cDNA synthesized from 10-fold serial-diluted human total RNA from 10 ng to 10 pg using AccuPower® RocketScript™ Cycle RT PreMix. (Gentaur, Cat. No K-2201) wase used as a template for PCR amplification. The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5min, 33 cycles of 95°C for 20 sec, 55°C for 20 sec and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
    Target : human GAPDH gene.
    Lane M : 100 bp DNA Ladder(Gentaur, Cat. No. D-1030)
    Lane 1 : 10 ng of human total cDNA
    Lane 2 : 1 ng of human total cDNA
    Lane 3 : 100 pg of human total cDNA
    Lane 4 : 10 pg of human total cDNA

    ProFi Taq fig3
    Figure 3. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 30 cycles of 95°C for 20 sec, 65°C for 20 sec and 68°C for 4 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
    Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
    Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
    Lane 1 : 2 kb fragment (human tumor protein p53 gene)
    Lane 2 : 3 kb fragment (human tumor protein p53 gene)
    Lane 3 : 4.5kb fragment (human DNA cross-link repair 1A gene)
    Lane 4 : 8 kb fragment (human hemoglobin epsilon 1 gene)

    ProFi Taq fig4
    Figure 4. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 32 cycles of 95°C for 20 sec and 68°C for 15 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol. Human genomic DNA was used as a template for PCR amplification.
    Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
    Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
    Lane 1 : 11 kb fragment
    Lane 2 : 13.5 kb fragment
    Lane 3 : 17.6 kb fragment
    Lane 4 : 21.4 kb fragment

    ProFi Taq fig5
    Figure 5. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 32 cycles of 95°C for 20 sec, 65°C for40 sec, and 68°C for 20 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol. Lambda DNA was used as a template for PCR amplification.
    Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
    Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
    Lane 1 : 15 kb fragment
    Lane 2 : 20 kb fragment
    Lane 3 : 25 kb fragment
    Lane 4 : 30 kb fragment

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