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2 × Taq Master Mix

价格 ￥147.00 （ 1-2周）
货号 UA070128
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产品介绍

产品介绍 评论(0)

产品规格

  • 性状

    Liquid

  • 缓冲体系

    20 mM Tris-HCl (pH 8.6 @ 25°C), 100 mM KCl, 3 mM MgCl2, 0.1% Tween® 20, 0.1% IGEPAL® CA-630, 10% Glycerol

  • 储存条件

    Store at -25 ~ -15℃ for 2 years

  • 文献引用

    1. Lawyer F C,Stoffel S,Saiki R K ,et al. Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus.[J].Journal of Biological Chemistry, 1989, 264.
    2. Eom S H,Steitz T A,Wang J M .Structure of Taq polymerase with DNA at the polymerase active site[J].Nature, 1996, 382(6588):278-281.

操作步骤

  • 1. After thawing, gently rotate and briefly centrifuge.
    2. Prepare the reaction system according to the table:

    Component

    Volume

    2 × Taq Master Mix

    25 μl

    Forward Primer (10 μM)

    2 μl

    Reverse Primer (10 μM)

    2 μl

    Template DNA*

    x μl

    RNase Free dH2O

    Up to 50 μl

    *Different templates have different optimal reaction concentrations, and the following table shows the recommended amount of template for the 50 μl reaction system:

    Template type

    Template quantity

    Animal and plant genomic DNA

    0.1 - 1 μg

    Escherichia coli genomic DNA

    10 - 100 ng

    cDNA

    1 - 5 μl (No more than 1/10 of the total volume of the PCR reaction)

    Plasmid DNA

    0.1 - 10 ng

    λ DNA

    0.5 - 10 ng

    3. Gently mix and briefly centrifuge.
    4. Prepare the PCR reaction system according to the table:

    Step

    Temperature

    Time

    Number of Cycles

    Initial Denaturation

    94°C

    3 minutes

    1 cycle

    Denaturation

    Annealing**

    Extension

    94°C

    50-60°C

    72°C

    30 seconds

    30 seconds

    1 min/kb

    25–35 cycles

    Final Extension

    72°C

    10 minutes

    1 cycle

    Soak

    4°C

    Indefinite

    1 cycle

    * The preconditioning condition is suitable for the vast majority of amplification reactions and can be adjusted according to the complexity of the template. If the template structure is complex, the preconditioning time can be extended to 5-10 minutes to improve the preconditioning effect.
    ** The annealing temperature needs to be adjusted according to the Tm value of the primer, generally set at 3 to 5 ℃ lower than the Tm value of the primer; for complex templates, the annealing temperature needs to be adjusted and the extension time extended to achieve efficient amplification.

注意事项

  • 1. It needs to be dissolved completely before use to prevent uneven ion concentration.

    2. The number of cycles should be selected according to the purpose of the experiment. If the number of cycles is too small, it will lead to insufficient amplification; if the number of cycles is too large, the amplification will increase, but the mutation rate will also increase, and non-specific amplification will occur.

    3. Set the appropriate annealing temperature according to the Tm value of the primer. If the annealing temperature is too low, it will cause non-specific amplification; if it is too high, it may not be amplified.

    4. Because Taq DNA Polymerase also has certain reaction activity at room temperature, the PCR reaction system should be prepared on ice and then placed in the PCR instrument for reaction. This can reduce non-specific amplification that occurs during the preparation of the reaction and help obtain high specificity amplification results.

  • 生物活性

    • Using 5ng λ DNA (48502bp) as the template, the amplification primers for the target fragments of 500bp to 6kb were designed. The amplification was carried out using UA 2 × Taq Master Mix and the 2 × Taq Mix after repeated freeze-thawing. The similar product of competitor company N was used as a control. The figure shows the results of the amplified, and the amplified range of the UA product is consistent with that of the competitive product N, and the amplified ability is stronger.
      Its amplification length can reach 6 kb, and it can efficiently amplify fragments of 4 kb and below. When stored at -20℃, the activity of UA products can remain stable after repeated freezing and thawing for more than 15 times.
      Marker 15K DNA；
      Lane 1: 500bp；
      Lane 2: 1000bp；
      Lane 3: 2000bp；
      Lane 4: 4000bp；
      Lane 5: 6000bp；

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