Table 2. Primers for amplifying and sequencing GPR143 genomic segments.
| Exon | Forward primer (5′- 3′) | Reverse primer (5′- 3′) | Product size (bp) | Annealing temperature(°C) |
|---|---|---|---|---|
| 1A |
CTCCTCCGCCCGCCCAAGCATCAC |
CCCAGGCAGCCGAGAAGGTC |
464 |
68 |
| 1B |
CCGCGCCTAGGGACCTTCTGCT |
AACCCGCGGGCCTCTCGTCCTCAC |
399 |
70 |
| 2 |
CTTTCTTCCTTTTCCCTCCTTGTC |
GTTTGCTGCTGCTGCGATTTG |
360 |
58 |
| 3 |
CACGTGCGGCTTCCTGAC |
TTGGCCTCTTATAAAAATGA |
385 |
56 |
| 4 |
GGGCTTTCCTCTGTGTACATTTTC |
CCCTGAGACAACGGCCTAACC |
334 |
60 |
| 5 |
GCATTTCCCTTTTTGTTCTCATCC |
AGGCCTGCACATTTTCATTTATTG |
406 |
58 |
| 6 |
TTGCTTCCTGCCCCTCTGG |
ACTTGCTCCCCTGTCCTCTGT |
400 |
60 |
| 7 |
TGCACCTGGCCCTCTTAGTTTC |
TCAGGAGGCCAAGACAGAGGAT |
441 |
60 |
| 8A |
AAACCAACCCACCAACCAGTCAAC |
GCATGCTCAGGGCTTCGTCA |
395 |
60 |
| 8B |
CCAGCCCAGGGATTTCTCTT |
ACCCCGCCATGCACAGGAC |
329 |
60 |
| 9 |
AGCTGATGACAAACCTGCTAG |
CCCTTTCTCCTATCCTAAAG |
330 |
58 |
| FAM5 |
CCTAGGGACCGGATGGGACAAC |
CTCGCCAACAGTTACACAGCTC |
487 |
60 |
| FAM6 |
GTCCCCTTTCTTCCCTTCTCTACT |
GTGGTGTTTGTCTTTGGTTTTGTG |
297 |
60 |
| SSCP-1 |
AGCCACGCAGCTCGTGCTGAGCTTCCAGCC |
CCCAGGCGCTGATCAGATTCCAACCCGCGG |
250 |
70 |
| SSCP-2 |
CATCCTCTTATCTTGACTTCC |
CCCAGAGAGCTTCCCTAGT |
224 |
58 |
| SSCP-3 | CATGTTCTCTTTACCTGCTGC | AGTGAGACCTTGTCTCTGAAG | 227 | 56 |
Listed are primer sequences, sizes of PCR products, and annealing temperature used for the amplification. Primers 1-9 were used to amplify and sequence the GPR143 genimic segments. Primers FAM5 and FAM6 were used to detect the deletion boundaries in families 5 and 6, respectively. Primers SSCP-1 to 3 were used to amplify exons 1, 5, and 7 for heterduplex-SSCP analysis.