Primers used in specific amplification of sodium bisulphite treated non-methylated and methylated FMR1 alleles
| Primer | 5′→3′ sequence | GenBank ID: nucleotides | Concentration | Amplicon size* |
|---|---|---|---|---|
| *n, number of trinucleotide repeats. | ||||
| bp, base pair; PCR, polymerase chain reaction. | ||||
| Non-methylated allele PCR (non-Met-PCR) | ||||
| Non-Met-F | AAACACTCAACTCCATTTCAATTTCACTTCCA | L29074: 13708→13739 | 0.2 μM | 168 bp+3n |
| Non-Met-R | GAGTTTGTTTTTGAGAGGTGGGTTGTGGGT | L29074: 13935→13906 | 0.2 μM | |
| Methylated allele PCR (Met-PCR) | ||||
| Met-F | CCGCCTCTAAACGAACGACGAACCGACG | L29074: 13748→13775 | 0.2 μM | 108 bp+3n |
| Met-R | GGTTGCGGGCGTTCGAGGTTTAGTCGTC | L29074: 13915→13888 | 0.2 μM | |
| Methylated allele triplet primed PCR (mTP-PCR) | ||||
| mTP-F | GCCGCTACCAAAAAACGTACGACAACGCG | L29074: 13806→13834 | 0.2 μM | Not applicable |
| mTP-R | TACCGATACGCATCCCAGTTTGTCAGC(TCG)8 | Not applicable | 0.02 μM | |
| Tail-R | TACCGATACGCATCCCAGTTTGTCAGC | Not applicable | 0.2 μM | |