Primestar hs dna polymerase with the gc buffer

DNA sequences for each subject were extracted for SNPs rs7412 and rs429358 from the APOE ε2/ε3/ε4 haplotype. APOE was genotyped using the standard Sanger sequencing method (Sangon, Shanghai, China) with the following primers: 5′-ACGCGGGCACGGCTGTCCAAGG-3′ (forward) and 5′-GGCGCTCGCGGATGGCGCTGA-3′ (reverse). APOE was amplified using the following conditions: 1 cycle of 98 °C for 10 s, 35 cycles of 72 °C for 5 s, 1 cycle of 72 °C for 5 min. PCR was performed in a final volume of 30 μl containing 10 pmol of forward and reverse primers, and 50 ng of genomic DNA template using PrimeSTAR HS DNA Polymerase with the GC Buffer (Takara Bio). In all, APOE genotype detection in 64 SCD and 70 controls were obtained. Due to the hemolysis of blood samples, we do not have the APOE genotype results of one SCD and three controls. When we considered the APOE effect in brain structure and behavior in SCD and controls, we excluded these four subjects.

DNA sequences for each subject were extracted for SNPs rs7412 and rs429358 from the APOE ε2/ε3/ε4 haplotype. APOE was genotyped using the standard Sanger sequencing method (Sangon, Shanghai, China) with the following primers: 5′-ACGCGGGCACGGCTGTCCAAGG-3′ (forward) and 5′-GGCGCTCGCGGATGGCGCTGA-3′ (reverse). APOE was amplified using the following conditions: 1 cycle of 98°C for 10 s, 35 cycles of 72°C for 5 s, and 1 cycle of 72°C for 5 min. PCR was performed in a final volume of 30 μl containing 10 pmol of forward and reverse primers, and 50 ng of genomic DNA template using PrimeSTAR HS DNA Polymerase with the GC Buffer (Takara Bio).