KAPA2G Robust HotStart PCR Kit

Description

• 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 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.
• Application: KAPA2G Robust HotStart® PCR Kit has been used in:PCR amplification of saliva samples[1]PCR amplification for direct sequencing[2]Nested PCR[3]Amplification of GC- or AT-rich templates[4]Templates containing common PCR inhibitors e.g. salts, urea, SDS, or ethanolCrude sample PCR e.g. buccal swabs, cultured mammalian, yeast, or bacterial cellsColony PCRRT-PCR[5]
• Biochem/physiol Actions: KAPA2G Robust HotStart® DNA Polymerase generated DNA are similar to fragments generated with wild-type Taq DNA polymerase. 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. It may be used in 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.
• Features and Benefits: Efficient amplification of GC- and AT-rich targetsRobust performance across a wide range of GC- and AT-rich templatesIncreased PCR success ratesImproved tolerance to common PCR inhibitorsEfficient amplification from crude samplesHigher yield and sensitivity per unit of enzymeUnrivalled performance in colony PCRHigher yields and improved consistency of PCR direct from E. coli and yeast cellsQuick Notes:KAPA2G Robust HotStart PCR Kits 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.
• Quality: Each batch of KAPA2G Robust HotStart DNA Polymerase is confirmed to contain <2% contaminating protein (Agilent Protein 230 Assay). KAPA2G Robust HotStart PCR Kits are subjected to stringent quality control tests, are free of contaminating exo- and endonuclease activity, and meet strict requirements with respect to DNA contamination levels.
• Preparation Note: Always ensure that the product has been fully thawed and mixed before use. Reagents may be stored at 4°C for short-term use (up to 1 month). Return to -20°C for long-term storage. Provided that all components have been handled carefully and not contaminated, the kit is not expected to be compromised if left (unintentionally) at room temperature for a short period of time (up to 3 days). Long-term storage at room temperature and 4°C is not recommended. Please note that reagents stored at temperatures above -20°C are more prone to degradation when contaminated during use, and therefore storage at such temperatures is at the user′s own risk.
• Other Notes: For Research Use Only. Not for use in diagnostic procedures.
• Legal Information: HOTSTART is a registered trademark of Molecular BioProducts, Inc.