2×HotStart HiFi PCR Master Mix with Dye

The 2×HotStart HiFi PCR Master Mix contains ABP HiFi DNA Polymerase, dNTP, Mg²⁺, and an optimized buffer system. The stabilizer and enhancer in the master mix ensure the reagent stability and optimize the amplification reaction. This pre-mixed formulation saves time and reduces contamination due to a reduced number of pipetting steps required for PCR set up. HotStart HiFi PCR Master Mix contains inert, non-toxic dyes to visualize PCR mixing step, and allows direct loading of PCR products on to gels for electrophoresis.

The ABP HiFi DNA Polymerase is a high-fidelity DNA polymerase that combines both amplification efficiency and fidelity. It possesses 5'→3' polymerase activity and 3'→5' exonuclease activity, and the amplified product is blunt-ended, suitable for fragment amplification of the seamless cloning kit and amplification of the second-generation sequencing library. The ABP HiFi DNA Polymerase has an extension rate of 15-30 s/kb, and its fidelity is 50 times higher than that of conventional Taq, overcoming the shortcoming of low amplification efficiency and slow amplification speed of ordinary Pfu enzyme, thus shortening the reaction time.   

This master mix has broad adaptability to PCR templates and strong resistance to inhibitors, and can be used for the amplification of DNA or cfDNA templates extracted from various samples such as FFPE, saliva, stool, urine, or blood.

Special Features

• Convenient, ready-to-use mix.
• Hot start.
• Generates blunt-ended PCR products.

Applications

• High-fidelity PCR.

General Protocol

1.  Thaw template DNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes.

2.   Prepare the following reaction mixture in a PCR tube on ice:
      

3.  Mix, and then briefly centrifuge the contents.

4.  Perform PCR using the recommended thermal cycling conditions outlined below:
     

5.   Analyze the amplification products by agarose gel electrophoresis.

     
    Fig 1. Taking human genomic DNA as templates, the target fragments of 0.6 kb, 1.0 kb, 2.6 kb, 3.0 kb, 4.0 kb, 5.1 kb, 6.2 kb, 7.1 kb, 8.5 kb, 10.6 kb, 17.8 kb, 20.3 kb, and 21.4 kb were amplified,  respectively.