DNA Methylation Validation Methods: a Coherent Review with Practical Comparison

Table 3 qMSP C_t values and primers’ efficiencies

Locus and used primer set	Average C_t of samples (n = 10)				C_t of standards		Efficiency
Locus and used primer set	MSP	± SD	BSP	± SD	MSP	BSP	MSP measured	MSP counted	BSP measured
M Met	23.18	0.52	22.88	0.27	24.01	22.70	96.47	94.57	83.00
IM Met	34.54	1.23	24.33	0.35	29.23	24.51	81.13	83.85	94.78
IM Unm	32.21	0.50	24.33	0.35	31.99	25.15	125.80	78.63	94.78
U Unm	37.36	0.80	23.04	0.20	37.95	23.40	65.93	61.66	90.52

M methylated locus, IM intermediately methylated locus, U unmethylated locus, Met primers for methylated DNA sequence, Unm primers for unmethylated DNA sequence, MSP methylation specific primers, BSP bisulfite specific primers (methylation independent), SD standard deviation.

ISSN: 1480-9222