Primers used for detecting NAT1*14 allele
Primer | Sequence 5′–3′ | Annealing temperature (°C) | Comment |
Sense N539 | 539TCC TAG AAG ACA GCA A CG ACC 559 | Bsa0I site in alleles with G560 | |
Antisense N1110b | 1110GGC CAT CTT TAA AAT ACA TTT T1089 | 54 | Specific for alleles NAT1*10 and NAT1*14 |
N714 | 714GTG AAG CCC ACC AAA CAG697 | 52 |
The numbers refer to the corresponding nucleotides in the coding region of NAT1. The underlined nucleotides have been mutated from the NAT1 sequence of the wild type to introduce a Bsa0I site (indicated by italics) in all alleles except NAT1*14 and others without a G at 560.18 The sense primer and N1110b as antisense primer were used with genomic DNA as template. The product was used as template with N539 with N714 as antisense primer (“nested” PCR). Primers N539 and N714 were used to amplify genomic DNA as template also as has been described.18