A 523 bp product was amplified using either HotStarTaq Plus DNA Polymerase with standard ramping times on standard cyclers (S) or the QIAGEN Fast Cycling PCR Kit with fast ramping times on fast cyclers (F). [A] Agarose gel analysis of the PCR product showed identical, highly specific amplification for both techniques. [B] Overall PCR time, comprising cycling time and ramping time, was calculated. The reduced PCR time on the fast cyclers was due mainly to the QIAGEN Fast Cycling PCR Kit allowing a significant reduction in cycling time. M: markers.
The master mix format and HotStarTaq Plus DNA Polymerase (contained in the master mix) ensure fast and easy PCR setup at room temperature.
Three different PCR assays (IL9, PKC, and AGRT2) were performed in duplicate using the QIAGEN Fast Cycling Kit (QIAGEN) and a fast-cycling PCR solution from another supplier (Supplier AII) according to the manufacturer's instructions. The QIAGEN Fast Cycling PCR Kit provided highly specific results for each assay, whereas results using Supplier AII were unpredictable with high drop-out rates and nonspecific results (e.g., primer dimers). M: marker.
The QIAGEN Fast Cycling PCR Kit drastically decreases the time required for highly specific hot-start PCR. Significant time savings can be made for all fragments <3.5 kb in length — calculations were made for a 500 bp fragment.
[A] The proprietary PCR buffer contains Q-Bond, a molecule that facilitates the reduction of the annealing step to 5 seconds. Q-Bond increases the affinity of Taq DNA polymerases for short single-stranded DNA fragments, reducing the time needed for successful primer annealing. [B] Without Q-Bond, the primer and polymerase bind sequentially to the template, increasing the duration of the annealing step.