The advent of PCR technology has transformed the clinical diagnostic laboratory. Typically, DNA used in PCR assays is extracted according to established procedures which are time consuming and increase the overall cost and turn-around time of these clinical assays. PCR testing directly from blood has not been routinely used with regular Taq DNA polymerases, due to the presence of multiple PCR inhibitors in whole blood. To counter the inhibition of heme, immunoglobin G (IgG), sodium citrate, heparin, or EDTA in whole blood, KAPA Biosystems has developed a special Taq DNA Polymerase by engineering Taq that can withstand higher concentrations of blood inhibitors.
KAPA Blood PCR Kits have been validated for the direct amplification of DNA fragments from whole blood collected in EDTA anticoagulant tubes, on FTA Elute Cards, Whatman 903 Specimen Collection Paper (“Guthrie cards”) or regular filter paper. The kit comes in two separate formats: PCR Mix A for fluorescent detection (e.g. Promega Power Plex 16) and Mix B for analysis by agarose gel electrophoresis and ethidium bromide staining. The kit components are very simple; the only difference is with a single tube: either with Mix A or Mix B, and a corresponding protocol. As per the protocol, reactions can be optimized using three difference final volumes; 50ul, 25ul and 10ul. We have optimized using 25ul; this reaction volume only requires 2.5ul of whole blood. This has really saved a lot of our starting sample; our previous protocol required 200ul of blood.
The protocol is very straightforward: just add 2.5 ul EDTA blood (heparin not suggested) into 12.5 ul of the Kapa Blood PCR mix with primer mix (10 ul), vortex, and your sample is ready to go into the thermal cycler for amplification.
We previously used a column extraction method which took about 50 minutes and cost nearly 5 dollars, but the Kapa Blood PCR Kit saves us time and each reaction does not cost more than more than a dollar. Last but not least, the kit includes Taq pol and the required buffers which means we need to add only our blood sample and primer pair. We mainly use this kit for detection of JAK-2 in genomic DNA from whole blood.
I found Kapa Blood PCR kit smarter, easier, faster, much cost efficient tool for working with little volume of blood.