Supplementary Table 1 – primer sequences used to amplify TCOF1 for sequencing or QF-PCR

Primers were tagged with M13 sequences: forward primer tag: gtaaaacgacggccagt; reverse primer tag: caggaaacagctatgac. Exon 23 was amplified in two overlapping fragments. Primers for 3’UTR were used to sequence MLPA probe binding site.

M13-tagged primers
Exon / Forward primer (5’>3’) / Reverse primer (5’>3’) / Product size (bp -including tags)
1 / GAAAGAGGAGCCGGAAGTG / AGTCGCAGTGGGAAGGCA / 426
2 / CAAGTCATGAGTTTGGGGAGA / GGGCTATGGGGACAGATCTTA / 283
3 / TGCGGGAAGGTCTGTCTAGT / GAAGCTGGAGGTAGACTCTA / 327
4 / AGGTGAGATGAAACCTGTTTG / CTATCCTTCTCTGACATGTA / 271
5 / TTGGGTTCAGATGCAAGTGG / GGCTGTCCTGGAGACAAACT / 419
6 / TTTGATGAGCAGCTGGTTTG / CTGAAGCCCCAGACAGGTT / 275
6A / AGCTTTATCAACTGCTGAAGC / GCTCATGGCAGAGTGCTCA / 428
7 / ATCAGTCAAGGTGGGGAGTG / AGAGTGGGAGAACAAGTCC / 461
8 / AGACCCCAGCCCCTTACTC / AAACAGGATGAGGGGAGAGC / 490
9 / AGTAGCCCCATGCCTAAACC / GAATGTGAGTGAGGGGAGGA / 493
10 / TTGCTCTCCTCCCCTCACT / GAGTCCCAGTGTGAGGTTGG / 407
11 / CCAACCTCACACTGGGACTC / TCAGAGTCCCCCATCTGTTC / 453
12 / ATCTGCCACACCACAGTCAG / AGGTGTCTCCAGGAGTCCAA / 491
13 / ACGTGGTCCTTTGGACTCCT / CCAGTCAAGCCCATCCTCA / 357
14 / GGGCTCCAGCTCCAGAGT / GCTGGGCTCAGTGACTGGT / 331
15 / GTTGTGCCATAGTGGGGAGT / AAGACAGGAGCTGGATGTGG / 360
16 / TAACACCTTTGCCACATCCA / ATTACAGGTGTGAGCCACCA / 404
16A / AGTGCCTGAGGGTCATTG / GCAAGATCTGCCCTCTCCT / 315
17 / GTGGACCCTTTGCCTTGTAA / ACTCAGCCAGTGTCCTGTCC / 407
18 / GTTATTGCCGCTGCTGAGGA / AGCACATGTGGGTAAGTATG / 315
19 / CAGTTTTGCCCCTTTGACTG / AACCAAGTGCAGAGGTCATG / 391
20 / TTCCTGGCCCTACTGAAGTG / TACAGGTGGGGAAACTGAGG / 485
21 / TGAGGGACCTGCAGAGAGAC / TGCCCAAGGCCTCTGTGAA / 345
22 / TCACAGCAGGCCATGACTCG / GGAGGACTGTTGCTACAGGA / 384
23 -5’ / CTTAGGTACCATAAGATCTG / GGTCTCCCGATAGCTTCC / 376
23 -3’ / GGAAAGAAAGAAGGTGGTGGA / CATGGGAGGAATGAGACCAG / 500
24 / GACCCCAGCACTTAGGATTA / TCTCCTTCCTTCCCCAACAC / 316
25 / AATTCACTAGTCCTCAGGAG / GGTCTGCCTGGCTCTCTG / 352
3’UTR / AGGACTCAGCCCAGTTCTCA / TATGAAGCGTCCTGTGTTGG / 384
Primers for QF-PCR
ex1(1) / CAGAGATGAGTAAAACGCAGACC / GGGACTTGGACGACTCTCT / 104
ex1(2) / GGGAGCTACTTCCCCTGATCT / GCCCACGAACGCTTACCT / 102
3’UTR(1) / GCATTGGAGATGCCAGATT / CACTGCTCGGCTAGGAAAT / 104
3’UTR(2) / TCTTGGAAGGGACTGTCTCG / TTCACTGCTCACTGCCTGTC / 100

PCRs for sequencing were set up using Megamix Blue (Microzone), 50-100ng DNA and 1mM each primer, and reactions were carried out in a Dyad PCR machine (Bio-Rad) using the following conditions: 95°C 10 mins, (95°C 30sec, 60°C 45sec, 72°C 30sec) x 32, 72°C 10mins. Amplification conditions for QF-PCR are given in the text.