Application

KAPA2G Fast HotStart^® ReadyMix^™ has been used for:Routine PCRFast PCRReverse transcription polymerase chain reaction (RT-PCR)Genotyping and fluorescent PCR

Biochem/physiol Actions

KAPA2G Fast HotStart^® DNA Polymerase generated DNA fragments may be used for routine downstream analyses or applications, including restriction enzyme digestion, cloning, and sequencing. 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.

Features and Benefits

Reduce cycling times up to 75% : Extension times as low as 1 sec per kilobase Broad coverage of both AT- and GC-rich targetsHigh speed, higher performance : Single copy sensitivity from complex targets Increased yieldsQuick Notes : KAPA2G Fast HotStart ReadyMix PCR Kits contain the engineered KAPA2G Fast HotStart DNA Polymerase, developed for fast PCR. Use 1 sec extension time for amplicons ﹤1 kb and 15 sec/kb for longer amplicons, and save 20–70% of total reaction time. No need for specialized instrumentation or PCR consumables. Optimized buffer system offers improved yields, specificity and sensitivity, facilitating efficient primer annealing across a wide range of primer lengths, GC contents, and melting temperatures. For high reaction efficiency, do not exceed 25 µL reaction volumes. KAPA2G Fast HotStart ReadyMix contains 1.5 mM MgCl₂ at 1X. KAPA2G Fast HotStart ReadyMix with dye includes two inert tracking dyes to allow direct loading onto agarose gels.Reaction products are 3’;-dA-tailed and may be cloned into TA cloning vectors.

General description

KAPA2G Fast DNA Polymerase is a second-generation (2G) enzyme engineered for higher processivity and speed, offering significantly faster extension rates than wild-type Taq polymerase. In addition to speed, KAPA2G Fast provides higher yields and sensitivity than competitor enzymes across a broad range of targets. KAPA2G Fast DNA Polymerase 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.KAPA2G Fast HotStart^® DNA Polymerase is supplied in a convenient 2X ReadyMix format, containing all reaction components except primers and template.

Legal Information

ReadyMix is a trademark of Sigma-Aldrich Co. LLC

HOTSTART is a registered trademark of Molecular BioProducts, Inc.

Other Notes

For Research Use Only. Not for use in diagnostic procedures.

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.

Quality

Each batch of KAPA2G Fast DNA Polymerase is confirmed to contain ﹤2% contaminating protein (Agilent Protein 230 Assay). KAPA2G Fast ReadyMix is subjected to stringent quality control tests, is free of contaminating exo- and endonuclease activity, and meets strict requirements with respect to DNA contamination levels.