MLPA probes used to determine CFHR1 and CFHR3 copy number
| Probe no . | Gene, exon . | Ligation site . | Partial sequence (20 nt adjacent to ligation site) . |
|---|---|---|---|
| 1 | CFH, exon 1 | 151-152 | TGCTACACAA-ATAGCCCATA |
| 2 | CFH, exon 2 | 226-227 | GGTCTGACCA-AACATATCCA |
| 3 | CFH, exon 3 | 389-390 | TCCTTTTGGT-ACTTTTACCC |
| 4 | CFH, exon 4 | 490-491 | ATTACCGTGA-ATGTGACACA |
| 5 | CFH, exon 6 | 850-851 | AAAGAGGAGA-TGCTGTATGC |
| 6 | CFH, exon 9 | 1310-1311 | AATCAAAATC-ATGGAAGAAA |
| 7 | CFH, exon 11 | Intron 10, 5429 nt before exon 11 | TAGGTAGTCA-TATTTGGAAC |
| 8 | CFH, exon 12 | Intron 12, 1079 nt after exon 12 | TGGACACATT-ATGATTGAGT |
| 9 | CFH, exon 13 | 1925-1926 | AGTTGGACCT-AATTCCGTTC |
| 10 | CFH, exon 15 | Intron 15, 863 nt after exon 15 | AGCTGAGTGA-CATGAGGTAG |
| 11 | CFH, exon 18 | 2751-2752 | GGAACCATTA-ATTCATCCAG |
| 12 | CFH, exon 20 | 3206-3207 | TCTGCTATTA-ATGCATGTTA |
| 13 | CFH, exon 22 | Intron 22 | ACTCATCACA-GAGATTTTTC |
| 14 | CFH, exon 23 | 3680-3681 | ACCTGTTCTC-GAATAAAGCT |
| 15 | CFH, exon 23 | Intron 23, 5 nt after exon 23 | TCAATACATA-AATGCACCAA |
| 16 | CFH, exon 23 | Intron 23, 286 nt after exon 23 | CACTTATACA-TGCAATCCGT |
| 17 | CFH, exon 23 | Intron 23, 8775 nt after exon 23 | AGTCCGAGGT-AGAAAGGGAC |
| 18 | CFH, exon 23 | Intron 23, 21 283 nt after exon 23 and 1354 nt before CFHR3 exon 1 | GTGGTAATCT-TGGCTCTCAG |
| 19 | CFHR3, exon 1 | Intron 1 | AGGTAAGTTA-AAAGAGATCT |
| 20 | CFHR3, exon 2 | Intron 1 | CATTTTCTTG-TGGAATTACA |
| 21 | CFHR3, exon 3 | Intron 3 | CGGACGACAG-TCTCAGACTT |
| 22 | CFHR3, exon 4 | Intron 4 | GGGTTATATG-AATTCCTACA |
| 23 | CFHR3, exon 6 | Intron 5 | TTCCCCAACA-TCACAGCAGA |
| 24 | CFHR3, exon 6 | 1003-1002 reverse | TCCCTTCCCG-ACACACTGCT |
| 25 | CFHR1, exon 1 | Intron 1 | GGATAATTCA-ATTGAAATGG |
| 26 | CFHR1, exon 3 | Intron 3 | AGAGTTTCAG-GTCCATGTGT |
| 27 | CFHR1, exon 5 | Intron 5 | AATCTGTGAT-TATTTTGTTA |
| 28 | CFHR1, exon 6 | 945-946 | CCTGTTCTCA-AATAAAAGCTT |
| 29 | CFHR1, exon 6 | 1246-1247 | TTTTCCAAGT-TTTAATATGG |
| 30 | CFHR2, exon 1 | Intron 1 | AACTATGTCT-TGGAGTTTCG |
| 31 | CFHR2, exon 2 | Intron 2 | AGATCATAAA-CACTTGATAA |
| 32 | CFHR2, exon 3 | Intron 3 | AATACCTGTG-TGTGGTTTAT |
| 33 | CFHR2, exon 4 | 589-590 | ATGCTCCAGG-TTCATCAGTT |
| 34 | CFHR5, exon 1 | 136-137 | TGGGTATCCA-CTGTTGGGGG |
| 35 | CFHR5, exon 2 | 207-208 | TGAAGAAGAT-TATAACCCTT |
| 36 | CFHR5, exon 3 | 398-399 | CTTCAGGACT-AATACATCTG |
| Probe no . | Gene, exon . | Ligation site . | Partial sequence (20 nt adjacent to ligation site) . |
|---|---|---|---|
| 1 | CFH, exon 1 | 151-152 | TGCTACACAA-ATAGCCCATA |
| 2 | CFH, exon 2 | 226-227 | GGTCTGACCA-AACATATCCA |
| 3 | CFH, exon 3 | 389-390 | TCCTTTTGGT-ACTTTTACCC |
| 4 | CFH, exon 4 | 490-491 | ATTACCGTGA-ATGTGACACA |
| 5 | CFH, exon 6 | 850-851 | AAAGAGGAGA-TGCTGTATGC |
| 6 | CFH, exon 9 | 1310-1311 | AATCAAAATC-ATGGAAGAAA |
| 7 | CFH, exon 11 | Intron 10, 5429 nt before exon 11 | TAGGTAGTCA-TATTTGGAAC |
| 8 | CFH, exon 12 | Intron 12, 1079 nt after exon 12 | TGGACACATT-ATGATTGAGT |
| 9 | CFH, exon 13 | 1925-1926 | AGTTGGACCT-AATTCCGTTC |
| 10 | CFH, exon 15 | Intron 15, 863 nt after exon 15 | AGCTGAGTGA-CATGAGGTAG |
| 11 | CFH, exon 18 | 2751-2752 | GGAACCATTA-ATTCATCCAG |
| 12 | CFH, exon 20 | 3206-3207 | TCTGCTATTA-ATGCATGTTA |
| 13 | CFH, exon 22 | Intron 22 | ACTCATCACA-GAGATTTTTC |
| 14 | CFH, exon 23 | 3680-3681 | ACCTGTTCTC-GAATAAAGCT |
| 15 | CFH, exon 23 | Intron 23, 5 nt after exon 23 | TCAATACATA-AATGCACCAA |
| 16 | CFH, exon 23 | Intron 23, 286 nt after exon 23 | CACTTATACA-TGCAATCCGT |
| 17 | CFH, exon 23 | Intron 23, 8775 nt after exon 23 | AGTCCGAGGT-AGAAAGGGAC |
| 18 | CFH, exon 23 | Intron 23, 21 283 nt after exon 23 and 1354 nt before CFHR3 exon 1 | GTGGTAATCT-TGGCTCTCAG |
| 19 | CFHR3, exon 1 | Intron 1 | AGGTAAGTTA-AAAGAGATCT |
| 20 | CFHR3, exon 2 | Intron 1 | CATTTTCTTG-TGGAATTACA |
| 21 | CFHR3, exon 3 | Intron 3 | CGGACGACAG-TCTCAGACTT |
| 22 | CFHR3, exon 4 | Intron 4 | GGGTTATATG-AATTCCTACA |
| 23 | CFHR3, exon 6 | Intron 5 | TTCCCCAACA-TCACAGCAGA |
| 24 | CFHR3, exon 6 | 1003-1002 reverse | TCCCTTCCCG-ACACACTGCT |
| 25 | CFHR1, exon 1 | Intron 1 | GGATAATTCA-ATTGAAATGG |
| 26 | CFHR1, exon 3 | Intron 3 | AGAGTTTCAG-GTCCATGTGT |
| 27 | CFHR1, exon 5 | Intron 5 | AATCTGTGAT-TATTTTGTTA |
| 28 | CFHR1, exon 6 | 945-946 | CCTGTTCTCA-AATAAAAGCTT |
| 29 | CFHR1, exon 6 | 1246-1247 | TTTTCCAAGT-TTTAATATGG |
| 30 | CFHR2, exon 1 | Intron 1 | AACTATGTCT-TGGAGTTTCG |
| 31 | CFHR2, exon 2 | Intron 2 | AGATCATAAA-CACTTGATAA |
| 32 | CFHR2, exon 3 | Intron 3 | AATACCTGTG-TGTGGTTTAT |
| 33 | CFHR2, exon 4 | 589-590 | ATGCTCCAGG-TTCATCAGTT |
| 34 | CFHR5, exon 1 | 136-137 | TGGGTATCCA-CTGTTGGGGG |
| 35 | CFHR5, exon 2 | 207-208 | TGAAGAAGAT-TATAACCCTT |
| 36 | CFHR5, exon 3 | 398-399 | CTTCAGGACT-AATACATCTG |
Ligation sites refer to positions in Genbank reference sequences NM_000186.2 (CFH), NM_021023.3 (CFHR3), NM_002113.1 (CFHR1), NM_005666.2 (CFHR2), and NM_030787.1 (CFHR5). Probes 12 to 14 were designed in our laboratory. The remaining probes are from an MRC Holland (www.mlpa.com) kit (SALSA MLPA kit P236-A1 ARMD).