The KAPA Blood PCR Kit is designed for the amplifcation of DNA fragments directly from whole blood. The product is based on KAPA Blood DNA Polymerase, a second-generation enzyme derived through a process of molecular evolution and the frst DNA polymerase engineered specifcally for Whole Blood PCR. The enzyme is available in KAPA Blood PCR Mix A or B, two optimized, easy-to-use 2x formats containing all PCR components except primers and template (whole blood).

Applications:

KAPA Blood PCR Kits have been validated for the direct amplifcation of DNA fragments from whole blood collected in EDTA anticoagulant tubes, on FTA® Elute Cards, Whatman 903^® Specimen Collection Paper (“Guthrie cards”) or regular flter paper. Positive results have been obtained with blood stored in EDTA tubes at 4°C for >2 years, as well as with fresh and frozen blood. DNA fragments up to 3.5 kb (with a GC content <65%) have been amplifed successfully using KAPA Blood PCR Kits. The amplifcation of fragments with a high GC content (>65%) is possible in the presence of DMSO, but the maximum amplicon length is limited to ≤1 kb. Sample types: The product is not suitable for the amplifcation of DNA from blood collected in anticoagulant tubes containing heparin. It has not been validated for direct amplifcation from blood collected in other anticoagulant tubes (e.g. citrate, ACD). Positive results with other crude sample types, including buccal swabs and amniotic fuid, have been reported. KAPA2G Robust HotStart PCR kits are recommended for direct amplifcation from other crude sample types.

Species:

KAPA Blood PCR Kits have primarily been validated in Whole Blood PCR using human blood, but may also be used with blood from other species collected in a suitable manner. Positive results with blood from other mammals (mice, cats) and several bird species have been reported. Theoptimal concentration of blood in a KAPA B lood PCR must be determined empirically for each species. Blood from species with nucleated erythrocytes typically have to be pre-diluted to more closely approximate the DNA concentration present in the same volume of human blood.

Detection of pathogens:

DNA fragments derived from pathogens have been successfully amplifed from suitable blood samples using KAPA Blood PCR Kits. However, the product has not been validated for pathogen detection and is currently recommended for genetic testing. Whole Blood PCR with this product is a viable alternative to amplifcation of DNA purifed from blood (using crude extraction methods or DNA purifcation kits) for the following applications:

– Single amplicon or multiplex end-point PCR, using unlabelled or fuorescently labelled primers.
– SNP analysis, based on restriction endonuclease digestion of PCR products.
– Paternity testing using the PowerPlex^®16 System (Promega Corporation).

The product has not been validated for real-time/quantitative PCR applications using SYBR^® Green or related dyes or sequence-specifc probes (e.g. TaqMan®, Molecular Beacons).