Abstract
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.
Highlights
Isolation of Nucleic Acids from human peripheral blood samples serves as the most important source in studies pertaining to epidemiology, forensics and biomedical/molecular genetics
We have introduced a simple nonenzymatic salt precipitation method for DNA extraction which can be useful for genotyping as well as telomere length determination
The DNA obtained from buccal cells was successfully used for locus specific PCR amplification of minisatellites, microsatellites and single nucleotide polymorphisms
Summary
Isolation of Nucleic Acids from human peripheral blood samples serves as the most important source in studies pertaining to epidemiology, forensics and biomedical/molecular genetics. Several studies have been undertaken using buccal cells as a source of nucleic acids. DNA obtained from buccal cells has been used for genetic studies [1,2,3,4,5,6,7,8,9,10] and cancer associated changes in oral cavity [11]. Total RNA obtained from buccal cells and its usefulness for gene expression studies is comparatively novel. Telomere length determination from DNA from buccal cells is relatively new to population genetic and forensic medicine
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