KAPA2G ROBUST (100U)

Stock Code: 2795970
Manufacturer Part No: KK5023

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Descriptions

Application

KAPA2G Robust PCR Kit has been used for:Amplification of GC- or AT-rich templates and gene amplificationTemplates containing common PCR inhibitors e.g. salts, urea, SDS, or ethanolCrude sample PCR e.g. buccal swabs, cultured mammalian, yeast, or bacterial cellsColony PCRRoutine PCRPCR amplificationAvipoxvirus specific PCRSimplex PCRSemi-quantitative PCR

Biochem/physiol Actions

The fidelity of KAPA2G Robust is similar to that of wild-type Taq. It has an error rate of approximately 1 error per 1.7 x 10⁵ nucleotides incorporated. DNA fragments generated with KAPA2G Robust DNA Polymerase have the same characteristics as DNA fragments generated with wild-type Taq DNA polymerase, and may be used for restriction enzyme digestion, cloning, and sequencing applications. Like wild-type Taq, KAPA2G Robust has 5’;→3’; polymerase and 5’;→3’; exonuclease activities, but no 3’;→5’; exonuclease (proofreading) activity.

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 PCR Kits contain KAPA2G Robust 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 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 was evolved to solve inconsistent amplification across a broad range of amplicon types (GC- and AT-rich). KAPA2G Robust PCR Kit 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.The enzyme is supplied with three reaction buffers, and a proprietary enhancer for GC-rich PCR