2.3. Genotyping
Genomic DNA was extracted from the peripheral whole blood of patients using a modified version of the ammonium acetate salting-out method [36]. PCR of the G6PD and HBB exons and the adjacent intron regions was completed with the primers and PCR conditions described previously [2,37,38].An α thalassemia genotyping kit (Yaneng Bioscience, China)was used to detect common alpha thalassemiamutations in China. All PCR products were purified and sequenced in both the forward and reverse directions using the ABI PRISM 3730 DNA sequencer (Applied Biosystems) by BGI to identify mutations. Data was collected and analyzed using Chromas 2.33 and BLAST.