Analysis of CYP2D6 Allele Frequencies and Identification of Novel SNPs and Sequence Variations in Sardinians

Table 2

List of primers used in CYP2D6 Long PCR protocols. P–1780 was designed in a highly non-homologous CYP2D6/CYP2D7P/CYP2D8P 5′untranslated region (nucleotides –1780_–1758 according to AY545216 sequence [13]). P–1584_WT and P–1584_MUT are modification of primer –1584G>C (Table 3) by addiction of a 3′-dNTP complementary to wild-type (WT) nucleotide or SNP (MUT), to create an amplification allele-specific. The 5′ 10-mer tag was added to PCR primers, in order to improve PCR efficiency.

Nucleotides position	Name	5′ 10-mer tag	5′-sequence-3′	Direction
From −1780 to −1758	P–1780	ACGTTGGATG	GTCCTCCTGTCCTCAGTGGAT	Forward
From −1584 to −1559	P–1584_WT	ACGTTGGATG	CAGCCTGGACAACTTGGAAGAAGCC	Forward
From −1584 to −1559	P–1584_MUT	ACGTTGGATG	CAGCCTGGACAACTTGGAAGAAGCG	Forward
From 4706 to 4728	2D6–R [14]	ACGTTGGATG	ACTGAGCCCTGGGAGGTAGGTA	Reverse