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Genomic DNA extraction from non-viable PBMCs


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#1 Rashmi

Rashmi

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Posted 22 December 2012 - 06:50 PM

Hey everyone,

I am trying to extract genomic DNA from 1 million peripheral blood mononuclear cells. Cells were pelleted and were frozen without any kind of freezing media. So, I am guessing that most of them are non-viable.
I used a kit to extract DNA and I got around 1.5 to 2ug total DNA (10ng/ul in 200ul elution volume).

I was just wondering if this yield is ok or if I am losing a lot of DNA? The protocol says that total yield from viable PBMCs should be around 6-8ug. The extracted DNA has a 260/280 ratio of 1.87 and 260/230 ratio of 0.97.
I want to use the extraction DNA for a taqman open array.

I will really appreciate if you guys can share your views.


Thanks,
Rashmi




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