KAPA2G Fast HotStart Genotyping Mix

Uses

KAPA2G Fast HotStart^? Genotyping Mix has been used in:

• Genomic DNA extraction and in the preparation of master mix for PCR
• Extraction and amplification from mouse tail, ear, and toe tissue
• Extraction and amplification of DNA from other animal tissues

General Description

KAPA2G Fast HotStart^? Mouse Genotyping Kits include KAPA Express Extract, a novel thermostable protease and buffer system that allows for the extraction of PCR-ready DNA from mouse tissue in as little as 15 minutes, and KAPA2G Fast Genotyping Mix with dye, containing a DNA polymerase engineered via directed evolution for high processivity and extreme speed. The combination of KAPA Express Extract and KAPA2G Fast Genotyping Mix allows for the reliable extraction and amplification of DNA fragments from mouse tissue in as little as 1 hour, as compared to ≥1 day with conventional protocols. KAPA2G Fast Genotyping Mix (2X) is a ready-to-use master mix containing all components for fast PCR with stabilizers, two inert dyes except primers and template.

Biochem/physiol Actions

KAPA2G Fast Genotyping Mix generated DNA fragments are 3′-dA-tailed and may be cloned into TA cloning vectors, or 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. Unlike existing protocols that rely on Proteinase K digestion, extractions using KAPA Express Extract are conveniently performed in a single tube, without the need for hazardous chemicals, or multiple wash steps.