Oragene™ DNA Self-Collection Kit From DNA Genotek

The collection of samples that will yield large amounts of good quality DNA can be difficult if blood samples are not used. Blood sampling is not always practical, requires skill and should be done in an appropriate place, such as a clinic or doctor’s office. The Oragene™ DNA Self-Collection Kit is a completely non-invasive collection method. Moreover, since saliva samples collected with Oragene™ are stable at room temperature for months, there is no need to quickly send samples to the laboratory to purify the DNA.

The Oragene™ kit can be used for self-collection by study subjects, including young and aged individuals. Donors simply have to collect 2 ml of saliva using the Oragene™ DNA Self-Collection Kit. The Oragene™ vial is then capped and mixed by inversion, allowing the DNA to be released and stabilized.

Once in the laboratory, DNA can be easily and rapidly extracted; no specialized kits or pieces of equipment are required. For extraction, the Oragene™ saliva samples need only to be heated, treated with the Oragene™ Purifier (included in the kit) and ethanol precipitated. All the researcher needs are tubes, ice, centrifuge and 95% ethanol. The step-by-step protocol is straightforward and the total time needed is not more than 2 hours. The 2 ml samples are incubated in a water bath for 1 hour at 50°C. A 500 µL aliquot (the residual can be stored at room temperature) is immediately removed from each sample and 20 µL (1/25th volume) of Oragene™ Purifier is added. Each tube is mixed well and incubated on ice for 10 minutes. The tubes are centrifuged at 15,000 × g for 5 minutes. The supernatant is removed and transferred to a fresh tube; an equal volume of room temperature 95% ethanol is added. The sample is mixed by inversion and allowed to stand 10 minutes at room temperature. The sample is centrifuged at 15,000 × g for 2 minutes and the supernatant is removed and discarded. The DNA pellet is dissolved in 100 µL of TE buffer and can be analyzed by agarose gel electrophoresis. In my experience, the resulting yield of DNA is 90-100 ug with an A₂₆₀/A₂₈₀ ratio >1.6.

I have successfully used the Oragene™ DNA Self-Collection Kit to obtain DNA from blood sampling scared people, and have used the DNA for SNP genotyping. The quantity and quality of the DNA is amazing and the results from downstream applications are of high quality.