A reduction in PCR cycling time is an attractive proposition for most PCR users, as it allows for faster turnaround times from sample to results and improved throughput without the need for additional thermocyclers. Using wild-type enzymes, fast PCR often leads to reduced sensitivity and higher failure rates when applied across a variety of primer-template combinations.
Kapa Biosystems has successfully employed its molecular evolution technology to engineer DNA polymerases specifically for fast PCR. Two highly processive enzymes, namely KAPA2G Fast and KAPA HiFi, offer true fast PCR, performed under standard reaction conditions (reagent concentrations) without loss in performance (yield and/or sensitivity). Shorter total cycling times are achieved, not by artificial reduction in denaturation and annealing times, but primarily because these fast polymerases require significantly shorter extension times that wild-type A- and B-family DNA polymerases. Both enzymes are available in antibody-based HotStart formulations.
KAPA2G Fast HotStart DNA Polymerase is the enzyme of choice for high-throughput, routine PCR. A total extension time of 1 sec per cycle is sufficient for amplicons up to 1 kb, whereas 15 sec/kb per cycle is recommended for longer amplicons. Any assay that yields satisfactory results with wild-type (hot start) Taq may be converted to a KAPA2G Fast HotStart assay, with the following advantages:
- Significantly (20 – 70%) reduced cycling times without sacrificing performance.
- No requirement for specialized PCR consumables or thermocyclers.
- No extensive reaction optimization required.
- A proprietary reaction buffer that facilitates primer annealing, thereby improving reaction efficiency.
- The availability of a ready-to-use 2x KAPA2G Fast HotStart ReadyMix.
- Ideally suited for fast, multiplex PCR.
KAPA HiFi HotStart DNA Polymerase is a unique B-family (proofreading) polymerase that does not only offer superior fidelity, but also significantly shorter extension times (15 – 30 sec/kb per cycle) than the 1 – 2 min/kb per cycle required for other proofreading enzymes. KAPA HiFi HotStart therefore offers:
- Significantly reduced cycling times for the high fidelity amplification of fragments up to 15 kb from genomic targets.
- Robust performance across a wide range of amplicon types.
Fast Real-Time PCR
KAPA SYBR® FAST and KAPA PROBE FAST qPCR Kits have been optimized for high-speed, real-time PCR without loss in performance.