FOLT PCR - A SIMPLE PCR PROTOCOL FOR AMPLIFYING DNA DIRECTLY FROM WHOLE-BLOOD
PANACCIO, M; GEORGESZ, M; Lew, AM
FoLT (formamide low temperature) PCR is a protocol for amplifying DNA directly from whole blood without any preparative steps. Up to 10% (vol/vol) whole blood can be added directly into the tube containing the PCR mixture. There is no need for transfers, centrifugations, preboiling or any preparative step. It involves the use of formamide (18% vol/vol) as well as reduced incubation temperatures (cycles of 85-degrees, 40-degrees, 60-degrees-C). The type of anti-coagulant used was critical: sodium heparin or EDTA being superior to lithium or fluoride heparin. Our studies indicate that FoLT PCR probably works by reducing the amount of protein coagulation and allowing more DNA template to be accessible for amplification. The sensitivity of FoLT PCR is such that a single copy gene from 5.5 nucleated cells in 1 mul of whole blood can be detected.