Figure 4.

Real-time PCR of whole blood containing samples. A 250-bp target was amplified from Lambda DNA, using 2 U of the BR Taq 22 mutant, Fast Start Taq or Jump Start Taq enzymes. Four 5-fold dilutions of DNA, starting with 10 ng (lanes 1–4), were used in reactions containing no blood or 5% and 10% human blood. Lanes M, DNA standard ladder. PCR was performed in a real time cycler using SYBR Green I as a fluorescent dye at concentration 32×. The amplified products were analyzed in 2% agarose gel stained with ethidium bromide (top row), along with the fluorescence detection of the amplification and the melting curves (middle and bottom rows, respectively.) The red, green, blue and yellow curves correspond to background subtracted fluorescence values obtained with the four DNA dilutions in decreasing order.