Cat.No.：A02051A
Price: ￥150
Specifications：
This product is a 2× ultra-high fidelity PCR premix specifically designed for high GC content templates. The premix system contains a newly modified generation of ultra-high fidelity DNA polymerase, dNTPs, GC enhancers, and an optimized reaction buffer. When used, only template DNA, primers, and deionized water need to be added to initiate the amplification reaction.
The core DNA polymerase has a fidelity 120 times higher than that of Taq DNA polymerase, and it also has extremely high amplification efficiency and wide template adaptability. This enzyme has 3′→5′ exonuclease correction activity, and the amplified products have blunt ends, which can stably and efficiently amplify difficult templates with a GC content of ≥70% and no limit on length. The unique stabilizers added to the premix can ensure that the amplification performance of the product remains unchanged after 25 repeated freeze-thaw cycles, and the stability of use is excellent.
High GC PCR premix solution (2×) dedicated GC buffer system: Optimized ion strength and additive ratio, effectively opening the secondary structure of high GC templates;
High success rate: Can still efficiently amplify templates with GC content as high as 75%;
Reduces non-specificity: Enhanced buffer, significantly reduces primer dimerization and non-specific bands;
Compatible: Suitable for conventional PCR instruments and rapid PCR programs.
| Cat. No. | Component | Specification | Quantity |
| B02051A | High GC PCR Master Mix (2×) | 1 mL | 1 |
| B02051A | High GC PCR Master Mix (2×) | 1 mL | 5 |
Amplification of high-GC DNA fragments.
When I conducted the gel electrophoresis test, my oligonucleotide did not show the correct length. Why was that?
The oligonucleotides should be detected on a polyacrylamide gel containing 7M urea, and should be loaded together with a 50% formamide solution to avoid compression and the formation of secondary structures. Oligonucleotides of the same length but with different compositions can produce different electrophoretic results. dC migrates the fastest, followed by dA and dT, and the slowest is dG. Oligonucleotides containing ribonucleic acid are prone to produce blurry electrophoretic bands and usually have problems with secondary structures.
The primers used at the beginning could have been used for PCR, but they became ineffective over time. Why is that?
Before conducting PCR, the primers should be aliquoted in the amount suitable for single use. The aliquoted primers should be heated at 94℃ for 1 minute. They should be rapidly cooled on ice before adding them to the PCR reaction mixture. Some primers may self-anneal or curl up.
