A rapid salting out method for DNA extraction from buccal swabs
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Supported by National Natural Science Foundation of China (30860319), and Technology R & D Program of Jiangxi Province (2009BSB09502).

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    Abstract:

    ObjectiveTo establish a rapid salting out method for extraction of genomic DNA from buccal swabs. MethodsBuccal 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 TE. The rs1042522 loci of TP53 gene and rs12910984 loci of CHRNA3 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. ResultsThe 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. ConclusionThe present salting out method is rapid, simple, and economical for DNA extraction from buccal swabs. The obtained genomic DNA is of high quality.

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History
  • Received:May 10,2011
  • Revised:October 19,2011
  • Adopted:November 08,2011
  • Online: December 20,2011
  • Published:
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