KAPA2G Robust HotStart PCR Kit

KAPA2G Robust HotStart PCR Kit

Application

KAPA2G Robust HotStart® PCR Kit has been used for:Amplification of GC- or AT-rich templatesTemplates containing common PCR inhibitors e.g. salts, urea, SDS, or ethanolCrude sample PCR e.g. buccal swabs, cultured mammalian, yeast, or bacterial cellsColony PCRqPCRNested polymerase chain reaction (PCR) amplificationAmplification of DNA targets in PCRDNA amplification using conventional PCRRoutine PCR

Biochem/physiol Actions

DNA fragments created with KAPA2G Robust HotStart® DNA Polymerase have the same characteristics as DNA fragments generated with wild-type Taq DNA polymerase. It can be used for routine downstream applications, such as restriction enzyme digestion, cloning, and sequencing. Like wild-type Taq, KAPA2G Robust HotStart has 5′→3′ polymerase and 5’;→3’; exonuclease activities, devoid of 3’;→5’; exonuclease (proofreading) activity. The fidelity of KAPA2G Robust HotStart is similar to wild-type Taq. It shows an error rate of approximately 1 error per 1.7 x 105 nucleotides incorporated.

Features and Benefits

Efficient amplification of GC- and AT-rich targets Robust performance across a wide range of GC- and AT-rich templates Increased PCR success ratesImproved tolerance to common PCR inhibitors Efficient amplification from crude samples Higher yield and sensitivity per unit of enzymeUnrivalled performance in colony PCR Higher yields and improved consistency of PCR direct from E. coli and yeast cellsQuick Notes: KAPA2G Robust HotStart® PCR Kits with dNTPs contain KAPA2G Robust HotStart DNA Polymerase, engineered for high processivity and inhibitor tolerance. Both purified genomic DNA and crude samples (e.g. colony PCR) can be used as template. Use 0.5 U of KAPA2G Robust HotStart DNA Polymerase per 25 µL reaction. More challenging PCRs (GCrich,crude sample) may require higher enzyme concentrations. Use 15 sec/kb extension time per cycle, and increase to 30-60 sec/kb for difficult amplicons or templates. KAPA2G Buffers contain 1.5 mM MgCl2 at 1X. Additional MgCl2 may be added using the 25 mM MgCl2 solution provided in the kit. KAPA2G Buffer A is optimized for high yield, specificity, and sensitivity. KAPA2G Buffer B is recommended for inhibitorcontaminated template samples. KAPA2G GC Buffer is recommended for GC-rich PCR (>70% GC). KAPA Enhancer 1 can be used with KAPA2G Buffer A or B, particularly for GC-rich assays. Reaction products are 3′-dA-tailed and may be cloned into TA cloning vectors.

General description

The second-generation KAPA2G Robust DNA Polymerase evolves to solve inconsistent amplification across a broad range of amplicon types (GC- and AT-rich). This product 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.

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