法医学研究ジャーナル

Usefulness of Nucleic Acids (DNA/RNA) from Buccal Cells Isolated from Mouthwashes using a Modified Method

Abstract

Divyalakshmi Saini and Birajalaxmi Das

Human buccal cells are an easy source of nucleic acids (DNA/RNA) other than human blood. We have extracted buccal cells from mouthwashes of 10 individuals and tested its usefulness for genotyping, telomere length determination and gene expression. Genomic DNA was extracted using a salt precipitation method whereas total RNA was extracted using Hipura total RNA extraction kit. The quantity and quality of DNA and RNA was found to be satisfactory. PCR amplification was achieved using locus specific primers and genotyping was performed for minisatellites, microsatellites and Single nucleotide polymorphisms (SNPs). Telomere length was determined from the genomic DNA of buccal cells obtained from mouthwashes. Telomere length obtained from the DNA isolated from buccal cells and peripheral blood mononuclear cells was compared. Real time quantitative PCR was successfully performed for GADD45A, G6PD, B-Actin and B2M. In conclusion, our method of DNA /RNA extraction can be considered as an alternative method of sample collection for forensic medicine.