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    Friday, 01 March 2013 11:06

    AccuPower TLA PCR PreMix

    Rate this item
    (1 Vote)

    AccuPower TLA PCR PreMix is a high fidelity PCR master mix that contains an easy to resuspend lyophilized mix of TLADNA Polymerase, dNTPs, reaction buffer, tracking dye, and patented stabilizer conveniently packaged in individual tubes or plates. Simply add template, primers and water. The TLA DNA Polymerase in the premix is a cloned and modified derivative isolated from Thermococcus onnurineus NA1 and features a proofreading activity, an enhanced processivity, and robust amplification efficiency compared to Pfu DNA polymerase or Vent DNA polymerase. These features allow high fidelity DNA amplification as well as long range PCR up to 20 kb.

    AccuPower TLA PreMix high fidelity PCR master mix is available with or without loading dye. When loading dye is present you can load it directly into your gel after PCR – no glycerol or glucose is required.

     

    Features and Benefits

    Ease of use: Just add template and primers and start your PCR, dNTPs, buffer and enzyme are provided
    Stability: Stable at room temperature for a month and for 2 years in a -20°C freezer
    High fidelity: For subcloning and other critical applications



    Specifications

    5' to 3' exonuclease activity: No
    3' to 5' exonuclease activity: Yes
    3’ – A Overhang: No
    Fragment Size: 20 kb


    Application

    Gene synthesis, PCR or primer extension requiring high fidelity, blunt end PCR cloning or mutagenesis, any amplification requiring high fidelity

    5antibodies CycleScript RT gentaur RT-PCR

    Figure 1. Comparison of long PCR amplification between AccuPower TLA PCR PreMix and 2 high-fidelity PCR enzymes. Reaction mixtures were prepared and PCR cycling conditions were performed following the protocol recommendations.

     

    Lane M1: 1 kb DNA Ladder (D-1040)
    Lane M2: Lambda/Hind lll Marker (D-1050)
    Lane 1, 8, 15: 2 kb PCR product
    Lane 2, 9, 16: 5 kb PCR product
    Lane 3, 10, 17: 8 kb PCR product
    Lane 4, 11, 18: 10 kb PCR product
    Lane 5, 12, 19: 12 kb PCR product
    Lane 6, 13, 20: 15 kb PCR product
    Lane 7, 14, 21: 20 kb PCR product

    6power elisa pcr gentaur taq PyroHotStart HotStart

    Figure 2. Comparison of amplification efficiency of AccuPower TLA PCR PreMix and other equivalent products PCR was carried out using AccuPower TLA PCR PreMix and 3 high-fidelity PCR enzymes from other companies. Parallel reactions were performed following the protocol recommendations, using 100 ng, 10 ng, 1 ng and 100 pg human genomic DNA. A 1 kb fragment of the human IR gene was amplified in 30 PCR cycles.

     

    M: 100 bp DNA Ladder (D-1030)
    Lane 1, 6, 11, 16: 100 ng of human genomic DNA
    Lane 2, 7, 12, 17: 10 ng of human genomic DNA
    Lane 3, 8, 13, 18: 1 ng of human genomic DNA
    Lane 4, 9, 14, 19: 100 pg of human genomic DNA
    Lane 5, 10, 15, 10: Negative control template

    7qPCR gentaur antibodies PreMix DNA Ligation PreMix

    Figure 3. . Comparison of detection sensitivity between AccuPower TLA PCR PreMix, Taq DNA polymerase and 2 high-fidelity PCR enzymes in amplification of various sizes. Reaction mixtures were prepared and PCR cycling conditions were performed following the protocol recommendations.

     

    M: 1 kb DNA Ladder (D-1040)
    Lane 1, 6, 11, 16: 2 kb PCR product
    Lane 2, 7, 12, 17: 4 kb PCR product
    Lane 3, 8, 13, 18: 5 kb PCR product
    Lane 4, 9, 14, 19: 7 kb PCR product
    Lane 5, 10, 15, 10: 8 kb PCR product

    8accupower RocketScript RT PreMix accu power elisa

    Figure 4. . A 2 kb Lambda DNA fragment was amplified according to the protocol recommendations using various extension times ranging from 20 sec to 2 min.

     

    M: 1 kb DNA Ladder (D-1040))
    Lane 1, 6, 11, 16: 20 seconds of extension time
    Lane 2, 7, 12, 17: 40 seconds of extension time
    Lane 3, 8, 13, 18: 1 minute of extension time
    Lane 4, 9, 14, 19: 1 minute 30 seconds of extension time
    Lane 5, 10, 15, 10: 2 minutes of extension time

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