Methylation-specific polymerase chain reaction (MSP): primer sequences and reaction conditions
Gene . | Forward primer: 5′-3′ . | Reverse primer: 5′-3′ . | Temperature, °C (no. cycles) . | PCR products, bp . | Reference no. . |
|---|---|---|---|---|---|
| SOCS1: MSP3′* | |||||
| M-MSP | TTC GCG TGT ATT TTT AGG TCG GTC (nt: 1081 - 1104) | CGA CAC AAC TCC TAC AAC GAC CG (nt: 1218 - 1240) | 63 (35) | 159 | Yoshikawa et al22 |
| U-MSP | TTA TGA GTA TTT GTG TGT ATT TTT AGG TTG GTT (nt: 1072 - 1104) | CGA CAC AAC TCC TAC AAC GAC CG (nt: 1218 - 1246) | 63 (35) | 174 | |
| SOCS1: MSP5′* | |||||
| M-MSP | GTT GTA GGA TGG GGT CGC GGT CGC (nt: 186 - 209) | CTA CTA ACC AAA CTA AAA TCC ACA (nt: 335 - 312) | 63 (35) | 149 | Nagai et al23 |
| U-MSP | GTT GTA GGA TGG GGT TGT GGT TGT (nt: 186 - 209) | CTA CTA ACC AAA CTA AAA TCC ACA (nt: 335 - 312) | 63 (35) | 149 | |
| SHP1† | |||||
| M-MSP | GAA CGT TAT TAT AGT ATA GCG TTC (nt: 6857 - 6880) | TCA CGC ATA CGA ACC CAA ACG (nt: 6995 - 7015) | 60 (40) | 158 | Oka et al29 |
| U-MSP | GTG AAT GTT ATT ATA GTA TAG TGT TTG G (nt: 6857 - 6882) | TTC ACA CAT ACA AAC CCA AAC AAT (nt: 6993 - 7015) | 59 (35) | 158 |
Gene . | Forward primer: 5′-3′ . | Reverse primer: 5′-3′ . | Temperature, °C (no. cycles) . | PCR products, bp . | Reference no. . |
|---|---|---|---|---|---|
| SOCS1: MSP3′* | |||||
| M-MSP | TTC GCG TGT ATT TTT AGG TCG GTC (nt: 1081 - 1104) | CGA CAC AAC TCC TAC AAC GAC CG (nt: 1218 - 1240) | 63 (35) | 159 | Yoshikawa et al22 |
| U-MSP | TTA TGA GTA TTT GTG TGT ATT TTT AGG TTG GTT (nt: 1072 - 1104) | CGA CAC AAC TCC TAC AAC GAC CG (nt: 1218 - 1246) | 63 (35) | 174 | |
| SOCS1: MSP5′* | |||||
| M-MSP | GTT GTA GGA TGG GGT CGC GGT CGC (nt: 186 - 209) | CTA CTA ACC AAA CTA AAA TCC ACA (nt: 335 - 312) | 63 (35) | 149 | Nagai et al23 |
| U-MSP | GTT GTA GGA TGG GGT TGT GGT TGT (nt: 186 - 209) | CTA CTA ACC AAA CTA AAA TCC ACA (nt: 335 - 312) | 63 (35) | 149 | |
| SHP1† | |||||
| M-MSP | GAA CGT TAT TAT AGT ATA GCG TTC (nt: 6857 - 6880) | TCA CGC ATA CGA ACC CAA ACG (nt: 6995 - 7015) | 60 (40) | 158 | Oka et al29 |
| U-MSP | GTG AAT GTT ATT ATA GTA TAG TGT TTG G (nt: 6857 - 6882) | TTC ACA CAT ACA AAC CCA AAC AAT (nt: 6993 - 7015) | 59 (35) | 158 |
Tm indicates annealing temperature; M-MSP, methylation-specific polymerase chain reaction for the methylated allele; U-MSP, methylation-specific polymerase chain reaction for the unmethylated allele; nt, nucleotide; and bp, base pairs.