Table 1. PCR and extension primer sequences
Mutation CD 41/42 (-CTTT) IVS2 654 (C → T) nt —28 (A → G) CD 17 (A → T) rs2187610
PCR primer 1 5′-ACGTTGGATGTAACAGCATCAGGAGTGGAC-3′ 5′-ACGTTGGATGTAACAGTGATAATTTCTGGG-3′ 5′-ACGTTGGATGTAGGGTTGGCCAATCTACTC-3′ 5′-ACGTTGGATGTCACCACCAACTTCATCCAC-3′ 5′-ACGTTGGATGATGCCATTTCATGGTTACC-3′
PCR primer 2 5′-ACGTTGGATGCTATTTTCCCACCCTTAGGC-3′ 5′-ACGTTGGATGGAAACCTCTTACATCAGTTAC-3′ 5′-ACGTTGGATGAGCAATAGATGGCTCTGCCC-3′ 5′-ACGTTGGATGTCAAACAGACACCATGGTGC-3′ 5′-ACGTTGGATGGAAGTGAGGCTACATCAAAC-3′
Standard protocol
Extension primer 5′-GATCCCCAAAGGACTCAA-3′ 5′-TGATAATTTCTGGGTTAAGG-3′ 5′-AGCCAGGGCTGGGCATA-3′ 5′-TTCATCCACGTTCACCT-3′ 5′-ACCTTTCATTTGTTCATTGTTTT-3′
Terminator mix* CGT AC AC CGT ACT
Expected molecular weight of extended nonmutant allele 6,088 6,475 5,558 5,345 7,225 (G allele)
Expected molecular weight of extended mutant allele 5,735 6,804 5,887 5,683 7,569 (C allele)
SABER protocol
Extension primer 5′-GATCCCCAAAGGACTCAA-3′ 5′-ATATGCAGAAATATTGCTATT-3′ 5′-GATGGCTCTGCCCTGACTT-3′ 5′-TTACTGCCCTGTGGGGC-3′ 5′-ACCTTTCATTTGTTCATTGTTTT-3′
Terminator ddCTP ddATP ddCTP ddTTP ddCTP
Expected molecular weight of nonextended primer 5,462 6,443 5,771 5,193 6,952
Expected molecular weight of extended allele 5,735 6,741 6,044 5,482 7,225 (G allele)
  • * CGT mix is ddCTP/ddGTP/ddTTP/dATP in which dd indicates the 2′,3′-dideoxynucleoside. Similarly, AC mix is ddATP/ddCTP/dGTP/dTTP