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 |