Abi prism 3130 3500 genetic analyzer

Genome was purified from whole blood by SRL (Tokyo, Japan), which is a private clinical laboratory test company. Polymerase chain reaction (PCR) amplification was performed using the primer sets and PCR kits (Supplementary Table S1) as per the manufacturer’s instructions. All coding regions, exon–intron boundaries, 5′ untranslated region (UTR), and 3′ UTR of AKR1A1 were examined by directly sequencing the PCR products using BigDye Terminator Cycle Sequencing Kit (Applied Biosystems, Foster City, CA, United States) and ABI PRISM 3130/3500 Genetic Analyzer (Applied Biosystems). We read both strands when an inserted or deleted nucleotide yielded dual signals derived from the wild-type (WT) and mutant strands. AKR1A1 sequence analysis was performed based on RefSeq NM_006066. Variant data we found are deposited in the NCBI dbSNP: NCBI_subsnp# are 2137544288 and 5316170415–5316170441.