Phusion® Blood Direct PCR Kit:
Superior performance even at high blood concentrations

Phusion Blood Direct PCR Kit delivers superior performance even at very high blood concentrations. Kits designed for direct blood PCR from three suppliers were compared by amplifying a 588 bp genomic DNA fragment with increasing blood concentration in the reaction mixture. PCR was performed according to suppliers’ instructions (35 cycles, except 40 cycles with the kit from Supplier 3) using Piko® Thermal Cycler and ultra-thin walled UTW® reaction vessels. Total protocol times indicated at bottom. + and - denote control reactions.
 

Direct PCR from blood of various vertebrates with
Phusion® Blood Direct PCR Kit

Amplification of a 237 bp DNA fragment from whole blood of 7 different vertebrates (5 % blood in the reaction). Piko Thermal Cycler and UTW reaction vessels were used in PCR.

 

Direct PCR from blood with different anticoagulants with
Phusion® Blood Direct PCR Kit

Storage temperatures of the blood samples:   1: Room temperature,   2: +4°C,   3: -20°C
Amplification of a 1 kb DNA fragment from human blood preserved with EDTA, citrate, or heparin (5 % blood in the reaction). Piko Thermal Cycler and UTW reaction vessels were used in PCR.
 

Direct PCR from blood stored on various cards with
Phusion® Blood Direct PCR Kit

1: Whatman 903 Card,   2: FTA Gene Card,   3: FTA Elute Card
Amplification of 1 kb, 3.8 kb, and 7.5 kb DNA fragments from human blood collected on Whatman 903®, FTA® Gene Saver or FTA® Elute Cards (1 mm punches in 50 µl PCR reactions. For the 7.5 kb fragment, FTA Gene Card punch was pretreated for 3 min at 50°C in H2O before adding it to the PCR reaction mixture). Piko Thermal Cycler and UTW reaction vessels were used in PCR.