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. 2010 Nov;12(6):797–807. doi: 10.2353/jmoldx.2010.090199

Figure 3.

Figure 3

Real-time MSP to amplify methylated sequences (placental) of the RASSF1A marker. A: Real-time MSP carried out on four first trimester CVS-maternal blood DNA pairs [N2 (46,XY), N3 (46,XY), N4 (46,XY), and N1 (46,XY)] after bisulfite conversion. The forward primer has been designed on CpG +157, +168, +170, and +177, while the reverse primer has been designed on CpG +229, +246, +248, and +250 (see Table 3). B: Real-time MSP to selectively amplify methylated sequences (fetal) of RASSF1A marker from maternal plasma. Reactions were performed on DNA extracted from first-trimester maternal plasma, CVS, and maternal blood cells after bisulfite conversion of two pregnancies (N5 and N3). Primer pairs are the same used in A. The quality and the amount of all templates have been checked by amplifying samples with a primer pair specific for both methylated and unmethylated templates (control real-time PCR). Upper panels indicate fluorescence curves, whereas lower panels represent electrophoretic analysis of real-time products. MW marker: molecular weight marker. CVS: chorionic villus sample.