A rapid salting out method for DNA extraction from buccal swabs / 第二军医大学学报

ABSTRACT

Objective To establish a rapid salting out method for extraction of genomic DNA from buccal swabs. Methods Buccal epithelial cells were digested with cell lysate solution and proteinase K solution. Then the proteins were removed by salting out and centrifugation and DNA was precipitated with isopropyl alcohol. Finally, the precipitations of DNA were washed with 70% ethanol and were resuspended in T. The rsl04252 loc o TP5 gene and rsl291098 loc o CHRNA gene were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. The samples with different genotypes were confirmed by direct sequencing analysis. Results The DNA yield of single buccal swab ranged from 0. 68 to 2. 56 μg; the D260/D280 value ranged from 1. 77 to 1. 94. After PCR amplification and enzyme digestion, two single nucleotide polymorphisms (SNPs) of 10 samples were clearly genotyped. The results of PCR-RFLP agreed well with the results of direct sequencing. Conclusion The present salting out method is rapid, simple, and economical for DNA extraction from buccal swabs. The obtained genomic DNA is of high quality.