Table 2 MSP primers and PCR conditions.
Gene | Forward primer (5′–3′) | Reverse primer (5′–3′) | Tm and cycles | Size in bp | Locus | Reference |
---|---|---|---|---|---|---|
p15 | ||||||
M‐MSP | TGAGGATTTCG CGACGCGTTC | CGTACAATAA CCGAACGAC CGATCG | 63°C/35 | 162 | 9p21 | 15 |
U‐MSP | TGAGGATTTTG TGATGTGTTT | CATACAATAA CCAAACAAC CAATCA | 162 | |||
p16 | ||||||
M‐MSP | TTA TTA GAG GGT GGG GCG GAT CGC | GAC CCC GAA CCG CGA CCG TAA | 65°C/35 | 150 | 9p21 | 15 |
U‐MSP | TTA TTA GAG GGT GGG GTG GAT TGT | CAA CCC CAA ACC ACA ACC ATA A | 151 | |||
p18 | ||||||
M‐MSP | TTATCGAATTG TTATTTTCGTT CG | CGTCTCGCCG AAAAAATAA TC | 64°C/35 | 93 | 1p32 | 15 |
U‐MSP | GGGTTATTGAA TTGTTATTTTT GTTTG | CATCCATCTC ACCAAAAAAA TAATC | 95 | |||
RB | ||||||
M‐MSP | GGGAGTTTCGC GGACGTGAC | ACGTCGAAAC ACGCCCCG | 66°C/35 | 152 | 13q14 | 15 |
U‐MSP | GGGAGTTTTGT GGATGTGAT | ACATCAAAAC ACACCCCA | 152 | |||
p21 | ||||||
M‐MSP | TTG GGC GCG GAT TCG TC | CTA AAC CGC CGA CCC GA | 62°C/35 | 100 | 6p21 | 25 |
U‐MSP | TTA GTT TTT TGT GGA GTT G | CTC AAC TCT AAA CCA CAA | 120 | |||
p27 | ||||||
M‐MSP | AAG AGG CGA GTT AGC GT | AAA ACG CCG CCG AAC GA | 66°C/35 | 195 | 12p13 | 25 |
U‐MSP | ATG GAA GAG GTG AGT TAG T | AAA ACC CCA ATT AAA ACA | 212 | |||
p57 | ||||||
M‐MSP | TCG GTT AGG TTT GAG CGA GC | TAC GTA TAC GAA AAA CGC GAC GAC | 62°C/35 | 137 | 11p15 | 26 |
U‐MSP | TTG GTT AGG TTT GAG TGA GTG A | TCT ACA TAT ACA AAA AAC ACA ACA A | 139 | — |
PCR, polymerase chain reaction; M, methylated; U, unmethylated; MSP, methylation‐specific PCR.