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1.1×S4 Fidelity PCR Mix (SF212)

1.1×S4 Fidelity PCR Mix (SF212)

It is a 1.1 × ready-to-use mixture for efficient PCR amplification, which has high fidelity and extremely fast extension speed.


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Product Description

It is a 1.1 × ready-to-use mixture for efficient PCR amplification, which has high fidelity and extremely fast extension speed.



Highlights


1) Simple operation: 1.1× ready-to-use premix, no need to add ddH2O, minimize the experimental steps.

2) High fidelity: The fidelity is 30 times that of wild-type Taq enzyme.

3) Extremely fast extension speed: 10~15 s/kb.

4) Ultra high heat resistance: The polymerase activity has no obvious change after heat treatment at 98℃ for 1 h.




Introduction


    This product is a 1.1× ready-to-use rapid PCR master mix containing S4 Fidelity DNA polymerase, dNTPs and an optimized buffer system. It can be amplified by adding primers and templates, and can effectively amplify DNA fragments within 5 kb. The system contains an electrophoresis indicator, which can be directly loaded for electrophoresis detection after the amplification is completed.

    The enzyme has 5'→3' polymerase activity and 3'→5' exonuclease activity, and the amplified product has a blunt end, which can be directly used for blunt end cloning.




Composition


Contents

Amount

1.1×S4 Fidelity PCR Mix  

5×1.125 mL



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Introduction

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Conventional PCR

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    One Step Rapid       Cloning

Uniclone One Step Seamless Cloning Kit

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Different from the traditional seamless cloning kit, the kit can easily achieve 2~6 fragments seamless cloning.


Q1：Why is there no product or a small amount of product in the final result?

A1：

    1)Template degradation or poor template purity. Use electrophoresis to check template quality and use high-quality templates.

    2)The template concentration is too low. Appropriately increase the template usage.

    3)The primers are not suitable. Optimize primer design.

    4)The annealing temperature is not suitable. Set the annealing temperature gradient and explore the appropriate annealing temperature.

    5)The number of cycles is too small. Appropriately increase 5~10 cycles.

    6)Insufficient extension time. For complex templates, the extension speed can be appropriately increased to 20~30 s/kb.

 

Q2：Why do non-specific bands or diffuse bands appear in amplification?

A2：

    1)Poor primer specificity. Optimize primers to avoid nonspecific bands and amplification of primer dimers.

    2)The primer concentration is too high. Decrease primer concentration.

    3)Template excess or poor purity. Reduce the amount of templates and use high-quality templates.

    4)The annealing temperature is too low. Appropriately increase the annealing temperature or use the Touchdown PCR program.

    5)Too many cycles. Reduce the number of cycles to 25~30 cycles.


Manual: Click to download

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