Background
Methods
Human subjects and genomic DNA
PCR and mutation screening
exon | primer name | sequence 5'> 3' | TA°C | reference |
---|---|---|---|---|
2 | PITX2-exon2for: PITX2-exon2rev: | tag tct cat ctg agc cct gc gcg att tgg ttc tga ttt cct | 60 | Ref: [25] this paper |
3 | PITX2-exon3bfor: PITX2-exon3brev: | ttg ctc ttt gtc cct ctt tct cct cgg agt gtc taa gtt caa gca gca | 60 | this paper |
4a | PITX2-exon4afor: PITX2-exon4arev: | ccg cct ctg gtt tta aga tg gca aag acc ccc ttc ttc tc | 60 | this paper |
4b | PITX2-exon4bfor: PITX2-exon4brev: | ctt gac act tct ctg tca gg aag cgg gaa tgt ctg cag g | 60/56/52* | Ref: [25] |
5 | PITX2-exon5for: PITX2-exon5rev: | cag ctc ttc cac ggc ttc t ttc tct cct ggt cta ctt gg | 60 | Ref: [25] |
6 | PITX2-exon6for: PITX2-exon6rev: | gta atc tgc act gtg gca tc agt ctt tca agg gcg gag tt | 65 | Ref: [25] |
Sequencing
Results and Discussion
patients (n = 96) | controls | |||
---|---|---|---|---|
type of variation: | specific variation | variant frequency (%) | number of controls | frequency (%) |
intronic/UTR variations:
| 2–40T>C (5'UTR exon 2) 2–18T>C (5'UTR exon 2) IVS2+7A>G (intron 2) IVS2-106C>A (intron 2) IVS3+11G>T (intron 3) IVS4a+11G (intron 4a) IVS4a-62C>A (intron 4a) | 10 (10.4%) 0 (0%) 1 (1.04%) 17 (17.7%) 1 (1.04%) 30 (31,25%) 1 (1.04%) | 100 100 100 100 100 100 100 | 12 (12%) 1 (1%) 0 (0%) 20 (20%) 0 (0%) 39 (39%) 0 (0%) |
silent mutations:
| 30G>C (S10S) (exon 2) 63C>T (A21A) (exon 4b) | 1 (1.04%) 1 (1.04%) | 100 100 | 0 (0%) 2 (2%) |
polymorphism within coding region:
| 30C>T (S27F) (exon 3) 204C>A (P65T) (exon 4b) | 0 (0%) 96 (100%) | 100 100 | 1 (1%) 100 (100%) |