Supp. Table S2.
Oligonucleotide primers used for PCR amplification of the novel missense mutations and polymorphisms
| Location | Forward (5′>3′)# | Reverse (5′>3′)# | Annealing Temperatur e (°C) | PCR product size (bp) | Restriction enzyme | |
|---|---|---|---|---|---|---|
| Mutation§ | ||||||
| Ex. 9 | c.953C>G | TTGGGTGATCCTTTTATTGTCA | CAACCCGCATCTCCCATGA* | 58 | 195 | Mnl I |
| Ex. 9 | c.967G>A | AGTTGCCTTATGGGAGCTGC* | CACTGGCAAATCTTGCTTAAAA | 60 | 229 | Pst I |
| Ex. 10 | c.1151T>C | GTTTGGATGAATCAGACTCAAAT | ACAAAAGTTTACCATGGTTCCA* | 56 | 246 | Bsr I |
| Ex. 13 | c.1468T>A | TTCTGTCCACATGAGATGAGC | CCAACATTGAAATCCACCTTAT* | 59 | 215 | Bstx I |
| Polymorphism§ | ||||||
| UTR 5′ | c.42GC | GCTTCCTGGCAACGCCGATC* | ACTCATGGCCCTCTTCCTTT | 62 | 309 | Taq I |
| Ex. 1 | c.61GC | GTCAGCATCAGCGGCCTCCT | ACTCATGGCCCTCTTCCTTT | 62 | 415 | Sma I |
| Ex. 1 | c.75CA | GTCAGCATCAGCGGCCTCCT | ACTCATGGCCCTCTTCCTTT | 62 | 415 | Hga I |
| Intr. 1 | c.195+34GT | GTCAGCATCAGCGGCCTCCT | ACTCATGGCCCTCTTCCTTT | 62 | 415 | Bbv I |
| Intr. 7 | c.752+56TC | ATCTTGGTCATAAATTCAACAGC | GAGAATGTAATCAAATGGGGAGA | 60 | 305 | Taq I |
Legend: Ex=exon, Intr=intron;
§
GALC gene GenBank-EM BL accession no. NM_000153.2 and no. NC_000014.8
*
primer modified so asto introduce a new restriction enzyme cleavage site according to Schwartz et al. (1991)