Table 1.
Primers and enzymes used for allele-specific analysis by PCRsa
| Region or gene | Primer or probe (sequence) | Temp (°C) | SNP B/C | Enzyme | Product size(s) (bp) |
|---|---|---|---|---|---|
| IG-DMR | IG-DMR207 (TACGGAGATGTGCTGTGGAC) | 62 | A/G | NcoI | B, 104, 95, 37 |
| IG-DMR442 (CTCGCTAGTTCACGGAGGTC) | C, 199, 37 | ||||
| Gtl2 DMR | Gtl2 1915F (TGAAACCTGTTGGGCG) | 61 | T/C | HaeII | B, 109, 70, 48, 17 |
| Gtl2 2158R (CAATGGGAGGGGTACAGATG) | C, 97, 70, 48, 17, 12 | ||||
| KvDMR CTCF BS1 | KvDMR 2701F (CCCACCGAAGTAATCCAAAA) | 62 | C/T | Hpy188I | B, uncut, 398 |
| KvDMR 3098R (TCAGCTAGGAAGGGATGAGG) | C, 292, 106 | ||||
| KvDMR CTCF BS2 | KvDMR 2258F (CTGAGAAGCCAAGTGGATCG) | 62 | A/G | MfeI/Tsp509I | B, 214, 82 |
| KvDMR 2553R (CCACCAGCCTCAGCATATTT) | C, uncut, 296 | ||||
| H19 | HI3 (CCTCAAGATGAAAGAAATGGT) | 55 | G/A | NA, real-time PCR | NA, real-time PCR |
| HE5 (AACACTTTATGATGGAACTGC) | |||||
| Probe Mut (CCACCTGTCGTCCATCTCC-FL) | |||||
| Probe Anc (RED640-TCTGAGGGCAACTGGGTGTGG-P) | |||||
| HE2 (TGATGGAGAGGACAGAAGGG) | 55 | G/A | Cac8I | B, 173, 62 | |
| HE4 (TTGATTCAGAACGAGACGGAC) | C, uncut, 235 | ||||
| Igf2 | Igf18 (ATCTGTGACCTCTTGAGCAGG) | 58 | Tsp509I | B, 180, 20 | |
| Igf20 (GGGTTGTTTAGAGCCAATCAA) | C, 165, 20, 15 | ||||
| Gtl2 | Gtl3 (CCAAAGCCATCATCTGGAATC) | 55 | T/? | SfcI | B, 250, 87 |
| Gtl4 (CAGCCCTGTGAGGTAGGAAC) | C, uncut, 337 | ||||
| Dlk1 | Dlk2up (CTGGCTTTCTTCCCGCTGGAC) | 54 | T/C | DraIII | B, 212, 104 |
| Dlk317dn (GACACAGCCAGGGGCAGTTA) | C, uncut, 316 |
a
SNP, single-nucleotide polymorphism; B, B6 maternal allele; C, paternal allele; NA, not applicable.