Table 1

Primer sequences and PCR reaction conditions

Primer set	Forward primer (5’–3’)	Reverse primer (5’–3’)	Product size (bp)	MgCl₂/Tm/cycles	Reference
(1) Methylation-specific PCR (MSP)
LPP/miR-28
M-MSP	CGC GGT TTA ATT ACG GGG TGT AC	CCA AAA AAA TCC GAA CAA AAA ACG	119	1.5 mM/56°C/35X	—
U-MSP	TTG TGG TTT AAT TAT GGG GTG TAT	CAA AAA AAT CCA AAC AAA AAA CA	119	2.0 mM/58°C/38X	—
(2) Quantitative reverse transcription PCR
LPP	GTG CAA TGT GTG TTC CAA GC	TGG CAT AAT AGG CTC CTT GC	211		40
GAPDH	ACC ACA GTC CAT GCC ATC ACT	TCC ACC ACC CTG TTG CTG TA	452		—

GAPDH, glyceraldehyde 3-phosphate dehydrogenase; MSP, methylation-specific PCR; M-MSP, MSP for the methylated allele; Tm, annealing temperature; U-MSP, MSP for the unmethylated allele.