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DNA Yield with Oragene

H.C. Birnboim
DNA Genotek, Ottawa, Ontario, Canada

Oragene gives higher DNA yields than other oral collection methods. The median yield of DNA from Oragene samples is 110 g. In comparison, buccal swabs may yield as little as 1.9 g of DNA.


Introduction
The number of studies collecting genomic DNA from a large number of individuals is increasing rapidly. Non-invasive methods and techniques that permit selfcollection are preferred because they increase compliance rates and reduce costs. For this reason, many large-scale studies now use buccal cells from saliva as the source of DNA. The two most common methods are scraping the inside of the cheek with a brush and swishing the mouth with a mouthwash. Oragene is a DNA self-collection kit that is non-invasive and intuitive to use. The donor rinses his or her mouth with water to clear food particles and then spits 2 mL of saliva into the Oragene vial. Once the vial is closed, DNA is released and stabilized at room temperature.

The amount of DNA recovered from the oral cavity can vary widely depending on the collection method (Figure 1). This technical bulletin reports the amount of DNA obtained from saliva using the Oragene kit.


Materials and Methods
Saliva samples were obtained from 208 donors. Collection and purification of DNA was carried out according to protocols supplied with the Oragene kit. DNA yield was determined by the highly specific Fluorescence/DNase method (ref. 5). The F/D method quantitates DNA using SYBR Green I dye (Molecular Probes, Inc.) and DNase treatment.


Results
The DNA yield of Oragene/saliva samples from 208 donors is shown in Figure 2. The median amount of DNA was 110 g. The 25th percentile was 62 g and the 75th percentile was 158 g.


Conclusions
Oragene is a non-invasive DNA self-collection kit that can be used by untrained study subjects, including children and the elderly. The median DNA yield from Oragene/saliva samples is 110 g. This is significantly higher than other oral collection methods.


References
1. Cozier, Y., Palmer, J., and Rosenberg, L. (2003) Comparison of Methods for Collection of DNA Samples by Mail in the Black Womens Health Study. AEP. 14, 117-122.

2. Harty, L., Garcia-Closas, M., Rothman, N., Reid, Y., Tucker, M., and Hartge, P. (2000) Collection of buccal cell DNA using treated cards. Cancer Epidemiology, Biomarkers & Prevention. 9, 501506.

3. Montserrat, G., Egan, K., Abruzzo, J., Newcomb, P., Titus-Ernstoff, L., Franklin, T., et al. (2001) Collection of genomic DNA from adults in epidemiological studies by buccal cytobrush and mouthwash. Cancer Epidemiology, Biomarkers & Prevention. 10, 687-696.

4. Le Marchand, L., Lum-Jones, A., Saltzman, B., Visaya, V., Nomura, A., and Kolonel, L. (2001) Feasibility of collecting buccal cell DNA by mail in a cohort study. 10, 701-703.

5. DNA Genotek Technical Bulletin PR003 Sept 30, DNA Quantifi cation Using the Fluorescence/DNase (F/D) Assay. 2004.


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