I use many kits, and I suggest the DNA extraction of Iso-Helix. Otherwise, you can choose a lysis buffer from Macherey, and proceed with manual protocol.

Thanks Panagiotis, very speedy! Will check both the kit and the Macherey buffer.

You can use kits for example: Qiagen, or Norgen's Saliva DNA Isolation Kit

we use to invitrogen kit for bucal cells and it is very easy and fast to use (2 HOURS), but the concentration is about 25 ng/ul and you resupend the pellet in 100ul the H2O. for sample, is very low!!!!!

Hi Claire,

We use the Qiagen Blood mini kit for extracting DNA from buccal swabs. We do not use the whole swab. Only small pieces so it does not absorb all of the lysis buffer. We follow the protocol provided in the manual including 15 minutes to lyse the cells. Then elute the final DNA with the EB buffer provided in the kit heated to 37 C. Generally our applications work better in water. So, we follow this up with ethanol precipitation using sodium acetate. The final result is often a high yield (greater than 1000 ng) of clean DNA. Resuspend in 20 uls water for a minimum concentration of 50 ng per ul. Purification with the Qiagen kit takes about 30 minutes. Ethanol precipitation including the wash step takes about 60 minutes.

Thanks for the suggestions - I came across PrepGem Saliva kit (sold through VWR) which takes 25 min and you do in a thermal cycler. Anyone had any experience with it?

Thanks Doug, I think you and I must have been typing at the same time! We use Qiagen kits quite a lot in the lab so that sounds interesting.

There was a paper pertaining to this in 2007: "A simple and cost–effective protocol for DNA isolation from buccal epithelial cells". They have a detailed protocol, but I have never personally tried the method.

Oragene's kits are excellent as well, high DNA yield

Thanks @Jaqueline and @Vijay, I'll check out the paper you suggest. Vijay, I love your alternate method though - can't go wrong when you can actually see the DNA "materialise" before your very eyes! :)

Gerson, I saw the oragene kit - but don't you have to have taken the sample using their collection tubes?

yeah, they provide collection tubes and reagents. The main advantage is the yield and quality of the extracted DNA =D .We were having issues with DNA quality from regular swabs/ followed by column extraction, especially after long-term storage. Their kits seem to have solved this problem

You can use

http://www.danagen.es/en/products/nucleic-acid-purification/genomic-dna-purification/saliva-dna/

David

Try the kit os IsoHelix. It is very quick, reliable and chip....

Just to name few issues with the use of buccal swab as a method for obtaining DNA:

- high bacterial content that can accelerate degradation of DNA as well as results in low yields and quality DNA

- DNA extraction process requires high lysis and DNA binding volumes as well has multiple hands on and centrifugation steps for filtration based purification methods.

Recently a California based company, Mawi DNA Technologies has introduced an new concept for collecting, concentrating and stabilizing oral sample samples collected with standard cotton swabs at the point of collection, which address the issues mentioned above. The iSWAB non-invasive sample collection devices results in dsDNA with < 1% bacterial content with yields up to 30ug of high quality DNA. You can use any commercially available blood gDNA extraction kit with no further modifications.

This some of the copy i have found on their website, i hope this helps

The iSWAB-DNA device is equipped with a squeezing insert to enable the efficient removal of swab collected cells/sample in a proprietary stabilizing solution. The innovative design of iSWAB allows for thorough removal of the cells/sample captured by the swab and then release into a proprietary lysis and nucleic acid stabilizing buffer. The combination of the mechanistic release feature and proprietary lysis buffer allows for a high quantity of cells to be collected in a concentrated manner from single or several swabs in a small amount of lysis buffer while maintaining low bacterial contamination. The process enables long term preservation of the sample at the point of collection and is suitable for room temperature storage, transport and extraction. A non-specialized individual can collect multiple swabs in a single device which concentrates the sample without the need of any component of the swab to be included, significantly reducing processing time and costs at the lab. The iSWAB is a self collection device where the whole collection process does not exceed more than 5 minutes and can be performed by a professional or by non-professional (self collection) with equal ease and simplicity.