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-   -   DNA extraction from saliva (http://seqanswers.com/forums/showthread.php?t=17927)

coleenmac 02-23-2012 07:24 AM

DNA extraction from saliva
 
I'm trying to maximize yields of extracted DNA from saliva. I'm collecting saliva using the DNA Genotek Oragene kits. I have tried both the Genfind kit (although I know it is really designed for blood), as well as the DNA Genotek manual method using their prepIT L2P reagent. However, I'm not getting the expected yields from either.

Any suggestions on how to optimize these methods or even a different kit/protocol to try?

niceday 02-23-2012 07:48 AM

what levels are your yields?
We run DNA extraction from blood, saliva and bucal swabs. This is for both genotyping and sequencing.
If you can give me some more info I can possibly advise.

www.kbioscience.co.uk

billbiggs 02-23-2012 08:29 AM

Hi Coleen,

We always gotten best yields using the simple alcohol precipitation protocol from DNAgenotek…

Have heard that other groups have automated extraction on Qiagen system.

Hope that this helps.

Best,
bill

coleenmac 02-23-2012 10:06 AM

From one sample set I'm getting anywhere from 2.5ug from 500ul of sample to 9ug from 500ul of sample (across different samples). For the second sample set, I'm in the process of collecting more data on yields, but my first attempt with this sample set gave only 1.4ug from 500ul of sample.

billbiggs 02-23-2012 10:39 AM

That was the big issue with saliva...generally got way more than needed but the amount varied greatly from individual to individual...far more variable than whole blood...but also far less quantity

ECO 02-23-2012 12:20 PM

Don't forget it's not all human nucleic acid. :eek:


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