Cloning

Successful cloning and subsequent protein expression requires the selection of the most suitable DNA Polymerase or 2x master-mix. The two most common PCR cloning methods are TA cloning and blunt-end cloning, and the choice depends on the downstream strategy, nature of the vector, and the PCR enzyme. While TA cloning typically employs a thermostable Taq DNA Polymerase, TA cloning with Taq does have limitations such as the size of the insert and the inherent error-rate of this DNA Polymerase. Alternatively, blunt-end cloning involves the ligation of an insert into a linearized vector where both DNA fragments lack overhangs. Blunt-end inserts can be produced using high-fidelity DNA Polymerases with 3′→5′ exonuclease or proofreading activity. This proofreading activity improves the sequence accuracy of the amplified products; however, limitations include lower ligation efficiencies. Azura Genomics offers a range of PCR enzymes and 2x master-mix options to suit the strategies and complexities of cloning.

Cloning

Successful cloning and subsequent protein expression requires the selection of the most suitable DNA Polymerase or 2x master-mix. The two most common PCR cloning methods are TA cloning and blunt-end cloning, and the choice depends on the downstream strategy, nature of the vector, and the PCR enzyme. While TA cloning typically employs a thermostable Taq DNA Polymerase, TA cloning with Taq does have limitations such as the size of the insert and the inherent error-rate of this DNA Polymerase. Alternatively, blunt-end cloning involves the ligation of an insert into a linearized vector where both DNA fragments lack overhangs. Blunt-end inserts can be produced using high-fidelity DNA Polymerases with 3′→5′ exonuclease or proofreading activity. This proofreading activity improves the sequence accuracy of the amplified products; however, limitations include lower ligation efficiencies. Azura Genomics offers a range of PCR enzymes and 2x master-mix options to suit the strategies and complexities of cloning.

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  1. ExtremeTaq™ DNA Polymerase

    ExtremeTaq™ DNA Polymerase is an enhanced formulation containing Taq Polymerase, proprietary enhancers, hot-start antibodies, and a proof-reading component for trouble-free PCR reaction assembly and consistent performance. ExtremeTaq Polymerase delivers a unique balance of PCR sensitivity, high fidelity, and value.

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    Starting at $215.00

  2. ExtremeTaq™ HiFi Mix

    The Universal All-in-One PCR Mix for Everyday PCR

    ExtremeTaq™ HiFi Mix is an optimized 2x master-mix comprised of an enhanced Taq DNA Polymerase, optimized reaction buffer, MgCl2, and ultra-pure dNTPs. This versatile master-mix is ideally suited to all routine end-point PCR applications and challenging DNA targets such as complex GC-rich DNA and low-copy number samples.

    In order to confirm the ideal reagent for your application, please click here for our PCR Selection Guide.

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    Starting at $85.00

  3. ExtremeTaq™ HiFi Red Mix

    The Universal All-in-One PCR Mix for Everyday PCR.

    ExtremeTaq™ HiFi Red Mix is an optimized 2x master-mix comprised of an enhanced Taq DNA Polymerase, optimized reaction buffer, MgCl2, ultra-pure dNTPs, and inert red gel-loading dye. This versatile master-mix is ideally suited to all routine end-point PCR applications and challenging DNA targets such as complex GC-rich DNA and low-copy number samples.

    In order to confirm the ideal reagent for your application, please click here for our PCR Selection Guide.

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    Starting at $85.00

  4. TruFi™ II DNA Polymerase

    TruFi™ II DNA Polymerase is an ultra-high fidelity proofreading DNA polymerase exhibiting both 5'→ 3' DNA polymerase activity and 3'→ 5' proofreading exonuclease activity, enabling the correction of mismatches during the amplification process.

    In order to confirm the ideal reagent for your application, please click here for our PCR Selection Guide.

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    Starting at $135.00

  5. TruFi™ II Ultra 2x PCR Mix

    TruFi™ II Ultra 2x PCR Mix is an optimized 2x master-mix comprised of TruFi™ II DNA Polymerase, optimized reaction buffer, MgCl2, and ultra-pure dNTPs.

    In order to confirm the ideal reagent for your application, please click here for our PCR Selection Guide.

    Learn More

    Starting at $135.00

  6. TruFi™ II Ultra 2x PCR Red Mix

    TruFi™ II Ultra 2x PCR Red Mix is an optimized 2x master-mix comprised of TruFi™ II DNA Polymerase, optimized reaction buffer, MgCl2, ultra-pure dNTPs, and an inert red gel-loading dye.

    In order to confirm the ideal reagent for your application, please click here for our PCR Selection Guide.

    Learn More

    Starting at $135.00

  7. TruFi™ DNA Polymerase
    TruFi™ DNA Polymerase is a new generation, ultra high-fidelity DNA polymerase developed specifically to overcome the many disadvantages of conventional proof-reading enzymes such as poor sensitivity, significant inhibition in crude PCR assays, long extension times and the tedious optimization of buffer conditions and reaction parameters. Learn More

    Starting at $135.00

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