Table 2.
Polymerase chain reaction primers and conditions
Polymorphism | Primer sequence and concentration | Annealing temperature (°C) | MgCl2 (mM) |
ACE I/D | ACE1 (D allele specific): 5′-CATCCTTTCTCCCATTTCTC, 800 nM | 50 | 1.5 |
ACE2 (I allele specific): 5′-TGGGATTACAGGCGTGATACAG, 800 nM | |||
ACE3: 5′-ATTTCAGAGCTGGAATAAAATT, 240 nM | |||
AGT T174M | Forward inner primer (T allele specific): 5′-CTGGGGCTGTGAACACGCCCACCAACA, 100 nM | 55 | 2 |
Reverse inner primer (C allele specific): 5′-TGATAGCCAGGCCCAGCTGCTGCTGTCAAC, 100 nM | |||
Forward outer primer : 5′-TCTCCCCAACGGCTGTCTTTGGCACCCT, 10 nM | |||
Reverse outer primer: 5′-CACTGGCTCCCATCAGGGAGCAGCCAGT, 10 nM | |||
AGTR1 +1166 A>C | Forward inner primer (A allele specific): 5′-TCTGCAGCACTTCACTACCAAATGAACA, 1000 nM | 58 | 3.5 |
Reverse inner primer (C allele specific): 5′-TCTCCTTCAATTCTGAAAAGTAGCTGAG, 1000 nM | |||
Forward outer primer: 5′-GCCAAATCCCACTCAAACCTTTCAACAA, 100 nM | |||
Reverse outer primer: 5′-AAGCAGGCTAGGGAGATTGCATTTCTGT, 100 nM | |||
AGTR2 +1291G>T | Forward inner primer (T allele specific): 5′-TTGTTTTTCTGATATGTTTGTACAAGACTT, 600 nM | 54 | 3.5 |
Reverse inner primer (G allele specific): 5′-TCTAGGTTGTAAATATGTCTCACYAATAAC, 600 nM | |||
Outer forward primer: 5′-AATAAAATTTCCCCTAATCTTTTCTGAA, 200 nM | |||
Outer reverse primer: 5′-GGAAATCCTGATAAAAATGGATATAAAA, 200 nM | |||
AGTR2 +1297G>A | Forward inner primer (G allele specific): 5′-TCTGATATGTTTGTACAAGATTKTCAGTG, 600 nM | 56 | 3.5 |
Reverse inner primer (A allele specific): 5′-GTTACTTCTAGGTTGTAAATATGTCTCCCT, 600 nM | |||
Outer forward primer: 5′-AATAAAATTTCCCCTAATCTTTTCTGAA, 200 nM | |||
Outer reserve primer: 5′-GGAAATCCTGATAAAAATGGATATAAAA, 200 nM |
ACE, angiotensin converting enzyme; AGTR1, angiotensin II type 1 receptor; AGTR2, angiotensin II type 2 receptor; D, deletion; I, insertion.