Table 1.

Primer sequences and PCR reaction conditions for AATK/miR-1250

	Forward primer (5′ to 3′)	Reverse primer (5′ to 3′)	Tm/cycles/MgCl2	References
Methylation-specific PCR (MSP) for AATK/miR-1250
M-MSP	TCG GAT TGT ATT AGC GGA GTT TC	CGC CGC AAA TAC GAA ACG	57 °C/35×/2 mM	NA
U-MSP	TTT GGA TTG TAT TAG TGG AGT TTT	ACC ACC ACA AAT ACA AAA CA	55 °C/37×/2 mM	NA
Quantitative real-time reverse transcription-PCR
AATK	CGG GTT CAA GGA GTT TGA GA	GTG AGT GGC AGG ACG TAC AC	NA	NA
WDR1	CAG TGT CTG ACG GTG CAT AA	ACG TCC AGT TTC ACA ACT CC	NA	[23]
GAPDH	ACC ACA GTC CAT GCC ATC ACT	TCC ACC ACC CTG TTG CTG TA	NA	[22]
β-Actin	GGA CTT CGA GCA AGA GAT GG	AGC ACT GTG TTG GCG TAC AG	NA	[24]
Cloning of luciferase reporter constructs with SRBSs of miR-1250-5p
MAPK1	AAG GGC TAG CTT GTG TCC CTG TAT TAC CAA AA	AGG GTC GAC TCT GGG GAA CAT AAC GAA GG	55 °C/35×/2 mM	NA
WDR1	AAG GGC TAG CCC TTC CTT TTC TTT TTC AGT GC	AGG GTC GAC GCG CTC AAA GTG TTT TCA CA	55 °C/40×/2 mM	NA

M-MSP methylated MSP, U-MSP unmethylated MSP, Tm annealing temperature, SRBS seed region binding site