quality of DNA sample

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quality of DNA sample

Postby zetreque on March 16th, 2017, 9:00 pm 

Does the quality of DNA sample depend on where or what type of tissue was used?
Example: Hair, Finger Nail, Blood, Saliva.

Or is it just a matter of performing the right technique on the DNA extraction? Is it all going to be the same DNA?

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Re: quality of DNA sample

Postby BioWizard on March 16th, 2017, 10:46 pm 

Probably both. Getting DNA from nice live cells is going to be better (less fragmented) than getting it from dead/dying keratinized cells. And extraction technique will affect purity and yeild.
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Re: quality of DNA sample

Postby zetreque on March 16th, 2017, 10:50 pm 

BioWizard » Thu Mar 16, 2017 6:46 pm wrote:Probably both. Getting DNA from nice live cells is going to be better (less fragmented) than getting it from dead/dying keratinized cells. And extraction technique will affect purity and yeild.


I read a page tonight that made it sound like hair roots are 60-70% reliable for nuclear DNA.

Saliva sounded like the best bet for getting the most DNA which can be refined I assume to the highest quality.
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Re: quality of DNA sample

Postby Braininvat on March 16th, 2017, 10:53 pm 

I've been wondering about that. My guess is less differentiated cells would be best, like epithelials. Blood is probably less good, since RBCs have no nuclei...you'd want to separate out white cells? Saliva or mouth swabs would be rich in epithelials.
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Re: quality of DNA sample

Postby zetreque on March 17th, 2017, 7:36 pm 

I know there are a few factors to this question but how many micrograms of DNA is needed to sequence it?
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Re: quality of DNA sample

Postby Braininvat on March 18th, 2017, 3:37 pm 

I answered this elsewhere, but can't find it.

It's still in my android clipboard, though. Wow, I've got many clips there. Here...

This is what New England Biolabs recommend at their website:
For low complexity templates (i.e. plasmid, lambda, BAC DNA), use 1 pg–10 ng of DNA per 50 µl reaction
For higher complexity templates (i.e. genomic DNA), use 1 ng–1 µg of DNA per 50 µl reaction
You can see the range is quite broad. Usually for small genomes you would use lower concentrations, since you expect more copies of your target sequence per  ng.



maybe I should use this tablet more often. Can't even find old clipboard stuff on my windows 10 machine back at home.
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Re: quality of DNA sample

Postby zetreque on March 18th, 2017, 3:56 pm 

I thought about one microgram (ug) to sequence DNA, but that has it at .01 ug
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