TIANSeq HiFi Amplification Mix (dye+)
Library amplification PCR premix with high library yield, high fidelity and low base bias
Catalog number / packaging
No. of preps
NG219-02 5×1 ml
NG219-01 1 ml
StorageThis kit should be stored at-25~-15°C. Avoid repeated freezing and thawing. The shelf life is one year.
DescriptionTIANSeq HiFi Ampliﬁcation Mix is a new high fidelity PCR amplification premix, which is suitable for PCR amplification of high-throughput sequencing library and other PCR related cloning and detection. The Ultra HiFi DNA Polymerase in the pre-mixed solution is a new type of fast and high fidelity DNA polymerase developed by directed molecular evolution technology, which enhances the affinity of DNA polymerase to templates. This enzyme has improved amplification speed and extension ability, and success rate of PCR and products yield is increased. The enzyme also has excellent 3'- 5' exonuclease activity (Proofreading activity), and its fidelity can reach the level of mainstream products in the market, with the authenticity in the process of gene and library amplification ensured. In addition, the DNA polymerase in the product has a HotStart function, which can effectively control non-specific amplification and enzyme activity loss under low temperature, thus ensuring specificity and stability of PCR amplification. A PCR reaction enhancement system is also integrated in the Mix, which not only improves the stability of the Mix, enhances the tolerance of polymerase to PCR reaction inhibitors, but also improves the adaptability of Ultra HiFi DNA Polymerase to templates with different GC contents, so that the product has wide sample universality and sample input range, and minimizes the base preference of PCR reaction.
Required ReagentsPrimer, Template
■ High-efficiency amplification: Ensure the conversion rate and reduce the amplification cycles.
■ Low Preference: Balanced amplification efficiency for DNA templates with different GC% contents.
■ High specificity: With HotStart property and strong specificity.
■ High fidelity: The fidelity is 50 times as high as Taq DNA polymerase.
■ High sensitivity: Template input can be as low as 1 pg.
DownstreamNGS library PCR amplification, 1st-generation sequencing PCR amplification, high fidelity cloning, SNP detection, site-specific mutation, etc.
Figure 1. The library enrichment of genomic DNA with different GC ratios (genomic input 10 ng, 8 cycles amplification) was carried out simultaneously by using TIANSeq HiFi Amplification Mix and HiFi enzyme from Supplier K and N, and the library yield was detected by Agilent 2100. The results showed that TIANSeq HiFi Amplification Mix has high library yield, with a strong universality for different GC contents, and the library enrichment performance was better than other suppliers.
Use TIANSeq HiFi Amplification Mix and HiFi enzyme specially used for NGS library amplification from Supplier K and N for library amplification of the same genomic DNA (genome input is 10 ng). After sequencing, analyze the coverage and duplication rate of the library.
Figure 2. Amplify genome libraries with different CG contents using TIANSeq HiFi Amplification Mix and the HIFi from Supplier K and N. The result shows that the uniformity of TIANSeq HiFi Amplification Mix amplification library is good and without GC preference, which is equivalent to the results of company K and slightly better than the products of company N. The results show that the coverage of TIANSeq HiFi Amplification Mix amplification library is high, the duplication rate met the requirements, and the amplification performance of the library is equivalent to that of competitors.