Description
General description
KAPA Taq DNA Polymerase is a single-subunit Taq DNA polymerase from the thermophilic bacterium Thermus aquaticus, purified from recombinant Escherichia coli. KAPA Taq ReadyMix™ (2X) is a ready-to-use cocktail containing all components for PCR, except primers and template. KAPA Taq ReadyMix is available with and without dye. The 2X ReadyMix with dye consists of two inert tracking dyes to permit direct loading of PCR products onto agarose gels for analysis by electrophoresis, without adding a DNA loading solution. KAPA Taq and KAPA Taq HotStart® DNA Polymerase have 5′→3′ polymerase and 5→′3′ exonuclease activities, but no 3′ → 5′ exonuclease (proofreading) activity. The enzyme has 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.
Application
KAPA Taq ReadyMix™ with dye has been used in:
high throughput PCR
the amplification of low copy DNA templates
multiplex PCR
specific amplification of complex templates
RT-PCR
nested PCR (nPCR)
amplifying exome 23 of anaplastic lymphoma kinase (ALK)
polymerase chain reaction (PCR) amplification
Biochem/physiol Actions
KAPA Taq DNA Polymerase has 5′→3′ polymerase and 5′→3′ exonuclease activity. It does not elicit a 3′→5′ exonuclease (proofreading) activity. This enzyme system has an error rate of approximately 1 error per 2.2 X 10⁵ nucleotides incorporated. PCR products created with KAPA Taq are A-tailed and are appropriate for cloning into TA cloning vectors.
Features and Benefits
Key Features:
High performance:
Improved sensitivity, specificity, and yields
Novel buffer formulation facilitates specific primer annealing, leading to higher yield of specific product
Quick Notes:
KAPA Taq ReadyMix™ can replace any commercial Taq DNA polymerase in an existing protocol. The annealing temperature may need to be optimized to account for differences in formulation.
The KAPA Taq PCR system is suitable for the amplification of fragments up to 3.5 kb from genomic DNA or 5 kb from less complex targets.
The 2X KAPA Taq ReadyMix with dye includes two inert tracking dyes, which allow loading of PCR products directly onto agarose gels for analysis.
KAPA Taq ReadyMixes contain 1.5 mM MgCl₂ and 0.2 mM of each dNTP (at 1X).
Preparation Note
Handling:
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.
Analysis Note
Each batch of KAPA Taq DNA Polymerase is confirmed to contain <2% contaminating protein (Agilent Protein 230 Assay). KAPA Taq ReadyMixes 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.
Other Notes
For Research Use Only. Not for use in diagnostic procedures.
Legal Information
HOTSTART is a registered trademark of Molecular BioProducts, Inc.
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
