Figure 2 | Laboratory Investigation

Figure 2

From: Meth-DOP-PCR: an assay for the methylation profiling of trace amounts of DNA extracted from bodily fluids

Figure 2

Methylation analysis of the TA-p73 gene in cell lines U937 (methylated, M) and HL60 (unmethylated, U) after distinct modification reactions conducted with decreasing amounts of genomic DNA. Five hundred (lane A) 250, (lane B) 125 (lane C), 62.5 (lane D), 31.2 (lane E), 15.6 (lane F) and 7.8 (lane G) ng of genomic DNA were modified with sodium bisulfite and diluted in 30 μl of water. Aliquots of 2 μl (theoretically corresponding to 33, 16.5, 8.25, 4.1, 2.05, 1.02 and 0.51 ng of the original DNA) were subjected to Meth-DOP-PCR (upper panel) or to MSP (lower panel). 1/25 of the Meth-DOP-PCR product (corresponding to 1.32, 0.66, 0.33, 0.16, 0.08, 0.04, 0.02 ng, of the original DNA) was utilized for the subsequent MSP with the TA-p73 set of primers. In parallel experiments, 2 μl of modified DNA were mock amplified by Meth-DOP-PCR in reactions not containing the enzymes (middle panel).

Back to article page