This application note describes protocols for amplifying DNA directly from blood samples stored on Whatman FTA and 903 Cards using the Phusion Blood Direct…
The Thermo Scientific Phusion Blood Direct PCR Kit enables the amplification of genomic DNA directly from blood samples stored on Whatman FTA and 903 Cards without the need for prior DNA extraction or purification. The kit utilizes a unique, engineered hot-start enzyme based on fusion protein technology that allows efficient amplification even in the presence of inhibitors found in blood and storage cards. When combined with the Thermo Scientific Piko Thermal Cycler and Piko UTW reaction vessels, PCR products can be amplified in as little as 30 minutes.
The document details specific reaction conditions and cycling protocols for 25 μL and 50 μL reactions. It presents results demonstrating the kit's ability to amplify various genomic DNA fragments (ranging from 237 bp to 7.5 kb) from human, mouse, pig, cat, dog, and bovine blood preserved on different card types. The note also outlines a simple washing protocol to remove inhibitors from FTA cards, allowing for successful amplification in smaller reaction volumes.
| Reaction volume (2x buffer) | 25 μL or 50 μL |
|---|---|
| Polymerase type | Phusion Blood DNA Polymerase |
| Compatible storage cards | Whatman FTA and 903 Cards |
| Minimum PCR time | 30 minutes |
| Compatible thermal cycler | Thermo Scientific Piko Thermal Cycler |
No, the kit enables DNA amplification directly from blood samples without prior DNA extraction or purification steps.
The recommended extension time is 15 seconds for amplicons ≤ 1 kb, and 30 seconds per kb for amplicons > 1 kb.
Yes, the kit is compatible with blood from various species including human, mouse, pig, cat, dog, and bovine.