Kapa Express Extract Blood-PCR Note
KAPA Express Extract is a novel thermostable protease and buffer system that allows for the extraction of PCR-ready DNA from various tissue types in as little as 15 minutes.
The KAPA Express Extract system is designed for optimal tissue lysis and sample preservation. Unlike protocols that rely on proteinase K digestion, DNA extractions with KAPA Express Extract are conveniently performed in a single tube, without the need for hazardous chemicals and multiple washing steps. This greatly reduces the risk of sample loss and contamination. Tissue lysis is performed in a standard thermocycler, heating block, or waterbath, after which the sample is centrifuged and the DNA-containing supernatant recovered.
Extracts may be used directly in PCR, without quantification or dilution. Depending on the tissue type, each extract typically yields a sufficient amount of template for 50–500 PCRs, and may be diluted in TE buffer for longterm storage at -20°C. DNA extracted with KAPA Express Extract is compatible with any PCR system. However, KAPA2G Robust HotStart ReadyMix is recommended for the rapid and reliable endpoint amplification of DNA extracted with KAPA Express Extract.
The ReadyMix contains the novel KAPA2G Robust HotStart polymerase, engineered for improved processivity and tolerance to common PCR inhibitors through a process of directed evolution. The 2X ReadyMix is a ready-to-use master mix containing all components for fast PCR, except primers and template. The combination of KAPA Express Extract and KAPA2G Robust HotStart ReadyMix Kits enables extraction and amplification of DNA fragments from crude tissues in ≤2 hours, as compared to ≥1 day with conventional protocols.
KAPA Express Extract is ideally suited for the extraction of PCR-ready DNA from, but not limited to, the following sample types:
• Human tissue (FFPE samples; blood collected in EDTA tubes or on collection cards; buccal swabs; hair follicles; forensic samples)
• Animal samples (ear or tail clippings; hair follicles; blood; bone marrow; dried or fresh tissue from mouse and other mammals)
• Fish tissues (fin punches; fresh tissue; cold and hot smoked canned samples; ethanol-preserved samples)
• Insects (crushed)
• Bird feathers (calamus fragments).