Description
General description
KAPA2G Fast Multiplex PCR Kits contain a second-generation (2G) enzyme derived through a process of directed evolution. KAPA2G FAST HotStart® DNA Polymerase is an antibody-mediated hot start formulation engineered for higher processivity and speed, offering significantly faster extension rates than wild-type Taq DNA polymerase. In addition to speed, KAPA2G Fast provides higher yields and sensitivity than competitor enzymes, allowing for more uniform multiplexed PCR.
KAPA2G Fast HotStart PCR Kits are designed for fast PCR, in which total reaction times are 20–70% shorter than those of conventional PCR assays performed with wild-type Taq DNA polymerase. This can be achieved without sacrificing reaction performance and does not require specialized PCR consumables or thermocyclers.
Application
KAPA2G Fast Multiplex Mix has been used for:
Typing of transgenic organisms
Amplification of microsatellites
Typing and detection of pathogens
Amplification of multiple DNA fragments for single nucleotide polymorphism (SNP) genotyping
Semi-quantitative PCR
3-plex PCR
Biochem/physiol Actions
DNA fragments generated with KAPA2G Fast DNA Polymerase have the same characteristics as DNA fragments generated with wild-type Taq DNA polymerase, and may be used for sequencing, restriction enzyme digestion and cloning. Like wild-type Taq, KAPA2G Fast has 5′→3′ polymerase and 5′→3′ exonuclease activities, but no 3′→5′ exonuclease (proofreading) activity. The fidelity of KAPA2G Fast is similar to that of wild-type Taq; it has an error rate of approximately 1 error per 1.7 x 10⁵ nucleotides incorporated. PCR products generated with KAPA2G Fast are 3′-dA-tailed and may be cloned into TA cloning vectors.
Features and Benefits
Improve sensitivity, specificity, and yields
Uniform representation of all amplicons
Successful multiplex PCR with difficult, GC-rich targets
Increase speed without compromising performance
60% reduction in cycling time
Extension times as low as 15 seconds
Quick Notes:
KAPA2G Fast Multiplex Mix contains the engineered KAPA2G Fast HotStart DNA Polymerase, for fastand efficient multiplex PCR.
The KAPA2G Fast Multiplex Mix contains a buffer optimized for multiplex PCR, with 0.2 mM of each dNTP and 3 mM MgCl₂ (at 1X).
Use 0.2 μM of each primer, and 10–100 ng of template DNA per reaction.
Anneal at 60°C for 30 seconds.
Perform extension for 15 sec, and increase for longer amplicons, and/or highly multiplexed reactions.
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 the ReadyMix has 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.
Analysis Note
Each batch of KAPA2G Fast HotStart DNA Polymerase is confirmed to contain <2% contaminating protein (AgilentProtein 230 Assay). KAPA2G Fast Multiplex 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 contaminationlevels.
Other Notes
For Research Use Only. Not for use in diagnostic procedures.
Legal Information
HOTSTART is a registered trademark of Molecular BioProducts, Inc.
