KAPA Taq HotStart

Description

• General description: KAPA Taq DNA Polymerase is the single-subunit Taq DNA polymerase of the thermophilic bacterium Thermus aquaticus, purified from recombinant Escherichia coli. KAPA Taq DNA Polymerase has 5’g3’ polymerase and 5’g3’ exonuclease activity, but no 3’g5’ exonuclease (proofreading) activity. The enzyme system has an error rate of approximately 1 error per 2.2 x 105 nucleotides incorporated. In the HotStart formulation, the enzyme is combined with a proprietary antibody that inactivates the enzyme until the first denaturation step. This prevents nonspecific amplification during reaction setup, increases sensitivity, and improves reaction efficiency. PCR products generated with KAPA Taq HotStart are A-tailed and may be cloned into TA cloning vectors. KAPA Taq HotStart Buffer is a uniquely formulated buffer to facilitate specific primer annealing. This translates to higher yields of specific product when compared to traditional Taq buffers, and improved amplification of GC- and AT-rich templates. However, KAPA Taq HotStart DNA Polymerase may be used in combination with any standard Taq buffer with a pH of 8.3 or higher.
• Application: KAPA Taq HotStart® has been used for:High throughput PCRAmplification of low copy DNA templates[1]Multiplex PCRSpecific amplification of complex templatesRT-PCRAquarium DNA extraction to generate marker gene libraries[2]
• Biochem/physiol Actions: KAPA Taq HotStart® DNA polymerase and KAPA Taq exhibit 5′→3′ polymerase and 5′→3′ exonuclease activities, but no 3′→5′ exonuclease (proofreading) activity. The enzyme shows an error rate of approximately 1 error per 2.2 x 105 nucleotides incorporated. In the hot start formulation, the KAPA Taq is combined with a proprietary antibody that inactivates the enzyme until the first denaturation step, eliminating spurious amplification products and increasing reaction efficiency and sensitivity. KAPA Taq hotstart buffer is formulated uniquely to facilitate specific primer annealing. This translates to higher yields of the specific product when compared to traditional Taq buffers. It also enhances the amplification of GC- and AT-rich templates. KAPA Taq hotstart DNA polymerase might be used in combination with any standard Taq buffer with a pH of 8.3 or higher
• Features and Benefits: High performanceImproved sensitivity, specificity, and yields.Novel buffer formulation facilitates specific primer annealing, leading to higher yield of specific product.Quick Notes:KAPA Taq HotStart® DNA Polymerase can replace any commercial hotstart Taq DNA polymerase in an existing protocol. The final MgCl2 concentration and annealing temperature may need to be optimized to account for differences in formulation.The KAPA Taq HotStart Buffer is a uniquelyformulated buffer offering improved specificity and sensitivity, and improved amplification of GC- and AT-rich templates.The KAPA Taq HotStart Buffer does not contain MgCl2; MgCl2 (25 mM) is supplied separately to allow greater flexibility during reaction setup.The KAPA Taq HotStart PCR Kit is suitable for the amplification of fragments up to 3.5 kb from genomic DNA or 5 kb from less complex targets.
• Quality: Each batch of KAPA Taq HotStart DNA Polymerase is confirmed to contain <2% contaminating protein (Agilent Protein 230 Assay). KAPA Taq 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.
• Other Notes: For Research Use Only. Not for use in diagnostic procedures.
• Legal Information: HOTSTART is a registered trademark of Molecular BioProducts, Inc.