Single Nucleotide and Structural Variants of CYP2D6 gene in Kinh Vietnamese population
1,2 Ha Hai Nguyen, 1Thuong Thi Huyen Ma, 1,2 Nhung Phuong Vu, 3 Quynh Bach Thi Nhu, 4Thang Vu Hong, 1,2Ton Dang Nguyen, 1,2 Hai Van Nong.
1Institute of Genome Research, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam.
2Graduate University of Science and Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam.
3Haiphong University of Medicine and Pharmacy, 72A Nguyen Binh Khiem, Ngo Quyen, Hai Phong
4Hanoi Medical University, 1 Ton That Tung, Dong Da, Hanoi, Vietnam.
Address for correspondence: Hai Van Nong. PhD, Institute of Genome Research, Vietnam Academy of
Science and Technology, 18 Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam. Tel: +84-904-4102458.
Email: [email protected] 1
2 3 4
5 6
7 8
9
10
11 12 13
14
SUPPLEMENTARY
Table 1. Primers sequence for PCR and sequencing
Primers used for amplification of CYP2D6 exons
Region Primer Sequence products sizeExpected
Promoter-Exon2
CYP2D6 ProF 5'- ATGTGGGCTGGGCTGGGAGCAGCC -3'
2080bp CYP2D6 E2R 5’- TCCCTAGTGCAGGTGGTTTC -3’
Exon3-Exon4
CYP2D6 E3F 5’- CAAGGTGGATGCACAAAGAG -3’
801bp CYP2D6 E4R 5’- GAATCTCTGACGTGGATAGGA -3’
Exon5-Exon8
CYP2D6 E5F 5’- GGCCAAGGACTCTGTACCTCCTA -3’
1889bp CYP2D6 E8R 5’- CAGCAGGGAGGTGAAGAAGA -3’
Exon8-Exon9
CYP2D6 E8F 5’- CAGAATGTTGGAGGACCCAACA -3’
853bp CYP2D6 E9R 5’- TGCTCAGCCTCAACGTACCCCT -3’
Primers used for sequencing
Region Primer Sequence
Promoter CYP2D6 ProR 5’- CCAGTGCTTCTAGCCCCATA -3’
Exon 1 CYP2D6 E1F 5'- ACTGGCAGCACAGTCAACAC -3' Exon 2 CYP2D6 E2F 5’- TCCCTAGTGCAGGTGGTTTC -3’
Exon 3 CYP2D6 E3F 5’- CAAGGTGGATGCACAAAGAG -3’
Exon 4 CYP2D6 E4F 5’- AAGAAGTCGCTGGAGCAGTG -3’
Exon 5 CYP2D6 E5F 5’- GGCCAAGGACTCTGTACCTCCTA -3’
Exon 6 CYP2D6 E6F 5’- GGCAGAGATGGAGGTGA -3’
Exon 7 CYP2D6 E7F 5’- CGTGAGCCCATCTGGGAAA -3’
Exon 8 CYP2D6 E8R 5’- CAGCAGGGAGGTGAAGAAGA -3’
Exon 9 CYP2D6 E9R 5’- TGCTCAGCCTCAACGTACCCCT -3’
Supplementary Figure 1.
15 16
17
18
19
20
21
22
Supplementary Figure 1. Confirmation of CYP2D6*5 allele by LR-PCR. Lane 1: 1kb plus DNA
ladder. Lane 2-3: samples homozygous for wild-type genotype. Lane 3-4: samples heterozygous for CYP2C6*5 allele. Using primer set reported from previous work, CYP2D6 deletion (*5) would result in 2
fragments containing 3.2 kb product indicating *5 allele (primers Dup and Dlow) and 5.1 kb product indicating wild-type allele (primers DPKup and DPKlow).
23
24 25 26 27 28
29
