Construction of plasmid DNA
Gene sequences for CYP2C19*2, CYP2C19*3, and CYP2C19*17 were searched at
https://www.ncbi.nlm.nih.gov/SNP and downloaded; the 621-bp CYP2C19 * 2
(rs4244285, c.681G > A) nucleic acid sequence was found,
and the mutation site is G → A. Similarly, an 897-bp CYP2C19 * 3
(rs4986893, c.636G > A) nucleic acid sequence was found,
with the mutation site G → A. The 897 -CYP2C19 * 17 (rs12248560, c.-806C
> T) nucleic acid sequence has the mutation site C → T.
Additional sequences, CYP2C9 (rs1057910), CYP2C19 * 4 (rs28399504),
CYP2C19 * 5 (rs56337013), CYP2C19 * 6 (rs72552267), CYP2C19 * 7
(rs72558186), and CYP2C19 * 8 (rs41291556), were designed for specific
detection. The designed CYP2C19 * 2, CYP2C19 *3, CYP2C19 *17 wild-type
and mutant sequences were synthesized by General Biology; the designed
specific sequences were synthesized by Tsingke Biotechnology. The PUC-57
vector with ampicillin resistance was used (see Table S1 for sequence).
Plasmids were diluted in 1X TE (pH 8.0), and the CYP2C19 * 2 wild-type
and CYP2C19 * 2 mutant plasmids were mixed together with 1:1 to prepare
a CYP2C19 * 2 heterozygous type; the heterozygous plasmids were mixed
together with 1:1 to prepare CYP2C19 * 2/3 heterozygous and CYP2C19 *
2/17 heterozygous, CYP2C19 * 3/17 heterozygous and CYP2C19 * 2/3/17
heterozygous plasmids.