Benefits of a universal annealing temperature for PCR


Primer annealing is a critical step in the polymerase chain reaction, or PCR. Suboptimal annealing can drastically reduce the yield and specificity of your PCR, and even cause it to fail. When it's not possible to design primers with similar melting temperatures, or Tm, the primer with the higher Tm could bind to unintended targets, while the primer with the lower Tm would have difficulty binding at an annealing temperature chosen for these primers. This can cause your PCR to fail. To mitigate this issue, the annealing temperature of each primer set should be optimized. But this can be a long and tedious process. To address the need for optimization, Thermo Fisher Scientific has developed a PCR buffer with isostabilizing components, which increase the stability of primer-template duplexes during annealing. Using PCR enzymes with this buffer, successful amplification can be achieved at 60 degrees Celsius, even with primers of different melting temperatures. Another benefit is that the enhanced stability of primer-template duplexes in this innovative buffer allows for a single PCR extension time with amplicons of different lengths. Typically, a long extension time would result in nonspecific amplification, but the isostabilizing components help achieve PCR specificity, even with a longer extension time. This universal annealing buffer has been developed with Invitrogen Platinum II Taq DNA polymerase, and Platinum SuperFi II DNA polymerase, and Platinum Direct PCR Universal Master Mix. One of a kind, this buffer enables high yield and specificity at 60 degrees Celsius, while also allowing for PCR of short and long amplicons at the same time and using the same protocol, instead of sequential runs. Only offered by Thermo Fisher Scientific, the universal annealing buffer of the Platinum DNA Polymerases helps save time and simplify PCR protocols by: - Reducing the need for optimization of primer annealing, - Enabling primer-template annealing at 60 degrees Celsius, and - Allowing co-cycling of different PCR targets To learn more about these PCR benefits, please visit

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