The second-generation KAPA2G Robust DNA Polymerase was evolved to solve – achieve consistent amplification across a broad range of amplicon types (both GC and AT-rich). The versatility and robustness of the polymerase allows for consolidation of PCR cycling protocols and reaction conditions while increasing success rates. The high performance of the KAPA2G Robust DNA Polymerase eliminates the need for multiple enzymes and protocols – standardize your PCRs with a single enzyme solution.
The KAPA2G Robust DNA Polymerase offers:
KK5004 | KAPA2G Robust DNA Polymerase with dNTPs (100 U) | 100 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1, dNTP Mix (10 mM) and extra MgCl2 (25 mM). |
KK5005 | KAPA2G Robust DNA Polymerase with dNTPs (250 U) | 250 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1, dNTP Mix (10 mM) and extra MgCl2 (25 mM). |
KK5023 | KAPA2G Robust DNA Polymerase (100 U) | 100 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1 and extra MgCl2 (25 mM). |
KK5024 | KAPA2G Robust DNA Polymerase (250 U) | 250 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1 and extra MgCl2 (25 mM). |
KK5515 | KAPA2G Robust HotStart DNA Polymerase (250 U) | 250 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1 and extra MgCl2 (25 mM). |
KK5516 | KAPA2G Robust HotStart DNA Polymerase with dNTPs (250 U) | 250 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1, 10 mM dNTP Mix and extra MgCl2 (25 mM). |
KK5517 | KAPA2G Robust HotStart DNA Polymerase (500 U) | 500 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1 and extra MgCl2 (25 mM). |
KK5518 | KAPA2G Robust HotStart DNA Polymerase with dNTPs (500 U) | 500 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1, 10 mM dNTP Mix and extra MgCl2 (25 mM). |
KK5522 | KAPA2G Robust HotStart DNA Polymerase (100 U) | 100 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1 and extra MgCl2 (25 mM). |
KK5525 | KAPA2G Robust HotStart DNA Polymerase (2,500 U) | 2500 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1 and extra MgCl2 (25 mM). |
KK5532 | KAPA2G Robust HotStart DNA Polymerase with dNTPs (100 U) | 100 units, 5 U/µL. Supplied with 5X KAPA2G Buffer A, 5X KAPA2G Buffer B, 5X KAPA2G GC Buffer (all with Mg2+ at a 1X conc. of 1.5 mM), 5X KAPA Enhancer 1, 10 mM dNTP Mix and extra MgCl2 (25 mM). |
KK5701 | KAPA2G Robust HotStart ReadyMix (100 rxn) | 100 x 25 µl reactions. Convenient 2X master mix containing KAPA dNTPs, reaction buffer, and Mg2+ at a 1X final conc. of 2.0 mM. Just add template and primers. |
KK5702 | KAPA2G Robust HotStart ReadyMix (500 rxn) | 500 x 25 µl reactions. Convenient 2X master mix containing KAPA dNTPs, reaction buffer, and Mg2+ at a 1X final conc. of 2.0 mM. Just add template and primers. |
KK5704 | KAPA2G Robust HotStart ReadyMix with dye (100 x 25 µL reactions) | 100 x 25 µl reactions. Convenient 2X master mix containing KAPA dNTPs, KAPA2G Robust HotStart DNA Polymerase, reaction buffer, Mg2+ at a 1X final conc. of 2.0 mM, and loading dye. Just add template and primers. |
KK5705 | KAPA2G Robust HotStart ReadyMix with dye (500 x 25 µL reactions) | 500 x 25 µl reactions. Convenient 2X master mix containing KAPA dNTPs, KAPA2G Robust HotStart DNA Polymerase, reaction buffer, Mg2+ at a 1X final conc. of 2.0 mM, and loading dye. Just add template and primers. |
The KAPA2G Robust DNA Polymerase is a highly robust and versatile second-generation enzyme derived through a process of molecular evolution. The novel amino acid mutations in KAPA2G Robust DNA Polymerase offer higher processivity and specific activity, which translates to robust performance across a wide range of GC- and AT-rich templates and amplicons, as well as improved tolerance to common PCR inhibitors.
In the HotStart formulation, the enzyme is combined with a proprietary antibody that inactivates the enzyme until the first denaturation step. This eliminates spurious amplification products resulting from non-specific priming events during reaction setup and initiation, and increases overall reaction efficiency.
KAPA2G Robust Kits supplied with KAPA2G Buffer A and KAPA2G Buffer B and the proprietary additive, KAPA Enhancer 1, offer extended optimization options for diverse and difficult templates. Kits also contain KAPA2G GC Buffer, a novel buffer formulated specifically for GC-rich templates and amplicons. Like wild-type Taq, KAPA2G Robust DNA Polymerase has 5’-3’ polymerase and exonuclease activities, but no 3’-5’ exonuclease (proofreading activity). The fidelity of KAPA2G Robust DNA Polymerase is similar to that of wild-type Taq.
KAPA2G Robust DNA Polymerase Kits, and HotStart formulations thereof, are recommended for all standard end-point PCR assays, particularly those in which wild-type Taq DNA polymerase does not perform satisfactorily. Kits are particularly suited for:
Amplicons generated with KAPA2G Robust DNA Polymerase are suitable for routine downstream applications, including restriction enzyme digestion, cloning and sequencing.
Half of each of the PCR products obtained with 72 of the 96 primer sets used in this study were electrophoresed in a 1% TBE-agarose gel. Amplicons were loaded in order of increasing GC content, with the lowest GC content (27%, blue) at the top left hand side and the highest GC content (84%, red) at the bottom right hand side of each composite gel image. Primers selected for this study had variable primer lengths, sequence composition, theoretical melting temperatures and other design features. Some primers contained 5′-tails for post-PCR sequencing using M13 or other standard sequencing primers. KAPA2G Robust HotStart ReadyMix reactions (25 µL) were performed as outlined in the User Guide. All reactions contained 25 ng human genomic DNA. 5% DMSO was included in all reactions (KAPA2G Robust and Taq) targeting amplicons with a GC content >70%.
Amplification of a 1.5 kb fragment from 1 pg plasmid DNA in the presence of four common PCR inhibitors using the KAPA2G Robust HotStart PCR kit and wild-type hot start Taq polymerase. All reactions contained 0.5 units of enzyme per 25 µL reaction. KAPA2G Robust HotStart Buffer B was used throughout, with the addition of KAPAEnhancer 1 for reactions containing SDS. Cycling was performed with an Eppendorf Mastercycler epgradient S, using a standard 3-step cycling profile (35 cycles) with an annealing temperature of 64ºC and 1.5 min extension time per cycle for all enzymes.
Amplification of a cloned 2.7 kb insert from four commonly used E. coli strains (DH5a, DH10B, JM109 or BL21) using KAPA2G Robust HotStart (top) or wild-type Taq (bottom). Colonies (grown on LB-agar + Amp plates) were either resuspended directly in individual PCR reactions (left) or first resuspended in PCR grade water and then added to PCR reaction mixes (middle). For overnight cultures (prepared in LB + Amp), 1 μL was added directly to the PCR mix (right).
Amplification of a 2.5 kb (left) or 1.6 kb (right) fragment from the GSH1 gene from three commonly used S. cerevisiae strains (BY4742, FY23 and W303) using KAPA2G Robust HotStart (top) or wild-type Taq (bottom). Colonies (from YPD-agar plates) or YPD overnight cultures were first lysed in 50 μL volumes with NaOH or Zymolase (as indicated).