Detection methods for VCAM1 genotype assays
| VCAM1 SNP . | Primers . | Annealing temperature . | Detection method . |
|---|---|---|---|
| VCAM1 promoter | F: TATTTCAGTGGGGACAAGGC | 63°C | Sequencing and BsrI digest |
| C-2021T | R: GACATCTGAAGTCCTACCTGC | ||
| VCAM1 promoter | F: AAGGACCTCTGGGTTACTTGTTT | 55°C | Sequencing |
| T–1599G | R: CTCCTCTTGGATACTGATGTGGCT | ||
| T–1592C | |||
| VCAM1 promoter | F: AGCCACATCAGTATCCAAGAGGAG | 61°C | Sequencing |
| T–1379C | R: GACCAGTTCTTGTTCATTGTTCATTGTTGTATC | ||
| A–1356G | |||
| A–1242T | |||
| VCAM1 promoter | F: GATACAACAATGAACAAGAACTGGTC | 52°C | Sequencing |
| G–1148A | R: CAATAACCAACTCTATGTTCCTTTTC | ||
| VCAM1 promoter | F: CAAGAGATTTGCCACTTCAGATG | 55°C | RsaI digest and sequencing |
| T–833C | R: AAAAGGGACACCATAACTTCTTAG | ||
| VCAM1 exon 1 | F: AGTGGAACTTGGCTGGGTG | 63°C | Sequencing |
| C–109T | R: TCACTACTATCGCAAAACTGACTG | ||
| G–54A | |||
| VCAM1 exon 5 | F: GTGTCCCAGAGAAACCATTTAC | 63°C | ApaLI digest (A1150G only) and sequencing |
| C953T | R: GAAAACCACTTACAGTAGAGCTCC | ||
| A1150G | |||
| VCAM1 exon 6 | F: CGTTTTTGCTTGCGATTTG | 55°C | Cac8I digest and sequencing |
| G1238C | R: CCAGTATCTTCAATGGTAGGGATG | ||
| VCAM1 exon 9 | F: TAGACATTAATTGCATCCATTTTG | 63°C | Sequencing |
| C2079T | R: ATTCAGGGAAGTCTGCCTCTC | ||
| A2146T | |||
| A2208G |
| VCAM1 SNP . | Primers . | Annealing temperature . | Detection method . |
|---|---|---|---|
| VCAM1 promoter | F: TATTTCAGTGGGGACAAGGC | 63°C | Sequencing and BsrI digest |
| C-2021T | R: GACATCTGAAGTCCTACCTGC | ||
| VCAM1 promoter | F: AAGGACCTCTGGGTTACTTGTTT | 55°C | Sequencing |
| T–1599G | R: CTCCTCTTGGATACTGATGTGGCT | ||
| T–1592C | |||
| VCAM1 promoter | F: AGCCACATCAGTATCCAAGAGGAG | 61°C | Sequencing |
| T–1379C | R: GACCAGTTCTTGTTCATTGTTCATTGTTGTATC | ||
| A–1356G | |||
| A–1242T | |||
| VCAM1 promoter | F: GATACAACAATGAACAAGAACTGGTC | 52°C | Sequencing |
| G–1148A | R: CAATAACCAACTCTATGTTCCTTTTC | ||
| VCAM1 promoter | F: CAAGAGATTTGCCACTTCAGATG | 55°C | RsaI digest and sequencing |
| T–833C | R: AAAAGGGACACCATAACTTCTTAG | ||
| VCAM1 exon 1 | F: AGTGGAACTTGGCTGGGTG | 63°C | Sequencing |
| C–109T | R: TCACTACTATCGCAAAACTGACTG | ||
| G–54A | |||
| VCAM1 exon 5 | F: GTGTCCCAGAGAAACCATTTAC | 63°C | ApaLI digest (A1150G only) and sequencing |
| C953T | R: GAAAACCACTTACAGTAGAGCTCC | ||
| A1150G | |||
| VCAM1 exon 6 | F: CGTTTTTGCTTGCGATTTG | 55°C | Cac8I digest and sequencing |
| G1238C | R: CCAGTATCTTCAATGGTAGGGATG | ||
| VCAM1 exon 9 | F: TAGACATTAATTGCATCCATTTTG | 63°C | Sequencing |
| C2079T | R: ATTCAGGGAAGTCTGCCTCTC | ||
| A2146T | |||
| A2208G |
PCR genotyping reactions were carried out as described under the following cycling conditions: initial denaturation for 10 minutes, followed by 40 cycles of 94°C ×30 seconds, indicated annealing temperature (see above) ×30 seconds, 72°C ×40 seconds, and final extension for 10 minutes. PCR products were then sequenced or digested with a genotype-specific restriction endonuclease. Restriction digests were incubated at 65°C (BsrI) or 37°C (ApaLI,Cac8I, and RsaI) for 4 hours, and then resolved on an agarose gel.