Uses
KAPA2G Robust HotStart
? ReadyMix
? has been used in:
- PCR
- Real time RT-PCR
- Nested Polymerase Chain Reaction (nPCR)
- Amplification of GC- or AT-rich templates
- Templates containing common PCR inhibitors e.g. salts, urea, SDS, or ethanol
- Crude sample PCR e.g. buccal swabs, cultured mammalian, yeast, or bacterial cells
- Colony PCR
The KAPA2G Robust HotStart ReadyMix may be combined with KAPA Express Extract for fast and efficient DNA extraction and PCR. For more information, refer to the application notes for:
- Blood PCR
- Mammalian DNA barcoding
- Fish DNA barcoding
- FFPE PCR
General Description
The second-generation KAPA2G Robust DNA Polymerase was evolved to solve inconsistent amplification across a broad range of amplicon types (GC- and AT-rich). KAPA2G Robust HotStart
? ReadyMix
? enables higher processivity and specific activity, which translates to robust PCR performance, high sensitivity, and improved tolerance to common inhibitors. The high performance of the KAPA2G Robust DNA Polymerase is ideally suited for challenging PCR applications and difficult samples, eliminating the need for optimization using multiple enzymes and protocols.
Biochem/physiol Actions
DNA fragments generated with KAPA2G Robust HotStart? DNA Polymerase have the same characteristics as DNA fragments generated with wild-type Taq DNA polymerase and may be used for routine downstream analyses or applications, including restriction enzyme digestion, cloning, and sequencing. Like wild-type Taq, KAPA2G Robust HotStart has 5′→3′ polymerase and 5′→3′ exonuclease activities, but no 3′→5′ exonuclease (proofreading) activity. The fidelity of KAPA2G Robust HotStart is similar to that of wild-type Taq; it has an error rate of approximately 1 error per 1.7 x 105 nucleotides incorporated. PCR products generated by KAPA2G Robust HotStart are 3′-dA-tailed and may be cloned into TA cloning vectors.