Sizes and primer sequences of PCR fragments for SSCP analysis of the promoter and the 3′UTR of the TAFI gene
Primer . | Primer sequences . | DNA position* (5′ to 3′) . | PCR size (bp) . | Annealing temperature (°C) . | Enzyme used before SSCP . |
---|---|---|---|---|---|
Promoter | |||||
T1a | CCTCATAAAGCAGTGCAAC | − 2676 | 296 | 58 | |
T1b | GTATGCTGTATTCTGGGTG | − 2380 | |||
T2a | CACCTCAACTGGACTATGT | − 2474 | 404 | 58 | RsaI |
T2b | TGTTCCCTTGCAGTTTAGC | − 2070 | |||
T3a | ATGGATCTGAGAAGCACC | − 2150 | 367 | 60 | TaqI |
T3b | TTTCCTCTCTGGGGAAAC | − 1783 | |||
T4a | CACCTGTAGACTTTTGC | − 1971 | 357 | 59 | |
T4b | CACTGAAGGAGAGAAAG | − 1614 | |||
T5a | TTTTCTTTCTCTCCTTCAGTG | − 1634 | 260 | 59 | |
T5b | ATCAATGAGACTGGTTATCCA | − 1374 | |||
T6a | ACTTCACCCAAAGAAGC | − 1467 | 282 | 59 | |
T6b | CCTGGCCAGAAGAAAAT | − 1185 | |||
T7a | ATTTTCTTCTGGCCAGG | − 1200 | 435 | 61 | Hinf1 |
T7b | CCTACATTCCCATTGGT | − 765 | |||
T8a | ACCAATGGGAATGTAGG | − 782 | 385 | 59 | MboI |
T8b | CCTTGCCTCTGACTTTT | − 397 | |||
T9a | TGGATTGGATTAGCCAG | − 476 | 322 | 59 | |
T9b | TTGGCTTAGTTAGGCTG | − 154 | |||
T10a | CCCAAGGAGTTACACAT | − 254 | 279 | 59 | |
T10b | AAGCTTCATCCCAACAG | − 25 | |||
3′ part of exon 10 + 3′UTR | |||||
3′a | CTTGGCATGTCATTAGG | +1268 | 612 | 59 | BamHI |
3′b | TGCGGCATTTGTTGACA | + 1880 |
Primer . | Primer sequences . | DNA position* (5′ to 3′) . | PCR size (bp) . | Annealing temperature (°C) . | Enzyme used before SSCP . |
---|---|---|---|---|---|
Promoter | |||||
T1a | CCTCATAAAGCAGTGCAAC | − 2676 | 296 | 58 | |
T1b | GTATGCTGTATTCTGGGTG | − 2380 | |||
T2a | CACCTCAACTGGACTATGT | − 2474 | 404 | 58 | RsaI |
T2b | TGTTCCCTTGCAGTTTAGC | − 2070 | |||
T3a | ATGGATCTGAGAAGCACC | − 2150 | 367 | 60 | TaqI |
T3b | TTTCCTCTCTGGGGAAAC | − 1783 | |||
T4a | CACCTGTAGACTTTTGC | − 1971 | 357 | 59 | |
T4b | CACTGAAGGAGAGAAAG | − 1614 | |||
T5a | TTTTCTTTCTCTCCTTCAGTG | − 1634 | 260 | 59 | |
T5b | ATCAATGAGACTGGTTATCCA | − 1374 | |||
T6a | ACTTCACCCAAAGAAGC | − 1467 | 282 | 59 | |
T6b | CCTGGCCAGAAGAAAAT | − 1185 | |||
T7a | ATTTTCTTCTGGCCAGG | − 1200 | 435 | 61 | Hinf1 |
T7b | CCTACATTCCCATTGGT | − 765 | |||
T8a | ACCAATGGGAATGTAGG | − 782 | 385 | 59 | MboI |
T8b | CCTTGCCTCTGACTTTT | − 397 | |||
T9a | TGGATTGGATTAGCCAG | − 476 | 322 | 59 | |
T9b | TTGGCTTAGTTAGGCTG | − 154 | |||
T10a | CCCAAGGAGTTACACAT | − 254 | 279 | 59 | |
T10b | AAGCTTCATCCCAACAG | − 25 | |||
3′ part of exon 10 + 3′UTR | |||||
3′a | CTTGGCATGTCATTAGG | +1268 | 612 | 59 | BamHI |
3′b | TGCGGCATTTGTTGACA | + 1880 |
DNA numbering of the TAFI promoter region is according to a proposed transcription start site (9). DNA numbering of the TAFI cDNA sequence is according to (13) and (9) for the end of the 3′UTR.