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Taq DNA polymerase

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    Product description
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    Product Description
    Taq DNA Polymerase is a thermostable recombinant DNA polymerase expressed by thermophilic Thermus aquaticus with a molecular weight of 94 kDa. It has 5'→3' polymerase activity and 5'→3' exonuclease activity, but no 3'→5' exonuclease activity. The amplified product has 3'-dA and can be directly used for TA cloning.
    Specification: 5 U/µL.
    Transportation and storage methods
    Ice pack transportation. Store the product at -20°C. The validity period is 2 years.
    Activity definition
    Using activated salmon sperm DNA as a template/primer, at 74ºC, within 30 min, the activity of ingesting 10 nmol of total nucleotides as an acid insoluble substance is defined as 1 activity unit (U).
    Quality Control
    Exonuclease residue detection: 20 μL reaction system, 10U of this product and 0.6 μg λ-Hind Ⅲ, incubated at 74ºC for 1 h, the DNA electrophoresis band has no change.
    Endonuclease residue detection: 20 μL reaction system, 10 U of this product and 0.6 μg Supercoiled pBR322 DNA, incubated at 74ºC for 1 h, the DNA electrophoresis band remained unchanged.
    Detection of E. coli residual DNA: Add 2 U of this product to a 50 μL system, and use sterile ddH2O as a template to amplify the E.coil 16s rDNA gene. After 35 cycles, the amplified products were subjected to 1% agarose gel electrophoresis, EB staining, and no amplified bands.
    Precautions
    1. For your safety and health, please wear lab coats and disposable gloves for operation.
    PCR reaction system (prepared on ice)
    Component Volume (μL) Final concentration
    ddH2O to 50 -
    10×Taq Buffer(Mg2+ Free) 5
    25 mM MgCl2 3 1.5 mM
    dNTP Mix (10 mM each) 1 μL 0.2 mM
    Template DNA optional -
    Primer 1 (10 μM) 2 μL 0.4 μM
    Primer 2 (10 μM) 2 μL 0.4 μM
    Taq DNA Polymerase (5 U/μL) 0.4 μL 0.04 U/μL
    PCR amplification program
    Cycle steps Temperature (ºC) Time Number of cycles
    Predenaturation 94 30 s~5 min 1
    Transsexual 94 30 s 35
    Annealing 50~60 30 s
    Extend 72 60 s/kb
    Final extension 72 10 min 1
    1. Pre-denaturation temperature and time: 94ºC is recommended. Recommended pre-denaturation time: 30 s for simple templates such as plasmid DNA; 3 minutes for complex templates such as cDNA and genomic DNA; 5-10 minutes for templates with high GC content.
    2. Annealing temperature and time: 60ºC is recommended, and a temperature gradient can be set up to find the optimal temperature for annealing the index object as needed. It is recommended to set the annealing time to 20 s, which can be adjusted within 10 to 30 s. Too long annealing time may cause the amplified product to appear diffuse on the gel.
    3. Amplification product: Please store the PCR amplification product at -20ºC to prevent DNA degradation.
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