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DNA extraction Method from Blood

DNA extraction Method from Blood - DNA Forum

DNA extraction Method from Blood - Discuss DNA, the molecule of hereditary. Topics include DNA structure, DNA replication, DNA function.


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Old 10-14-2009, 11:32 AM
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Default DNA extraction Method from Blood



I am working in the field of Diabetes genetics. Please let me know which method is best to extract DNA from Blood sample, Here in may lab they have some serum, Plasma and blood cell.it is possible to isolate DNA from them if some one any experience Please share.....
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Old 10-14-2009, 02:35 PM
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Default Re: DNA extraction Method from Blood

There will be DNA in all of the above materials, but at low concentration. Nearly all of the cells in the blood (in terms of total numbers) are DNA-free: both RBC's and platlets (the most numerous cells) are anucleate.

Your best bet, if its an option, is to start with whole blood. Spin it down to get the buffy coat (basically, concentrated WBCs) and use that as your source of DNA. The WBC's are the major source of DNA in blood - plamsa, RBCs, serum, etc, will only contain trace amounts.

Another option, if you're really stuck, is to use glassmilk to purify free DNA from the serum/plamsa. Spin down any particulates out of the plasma, mix 10ul glassmilk/ml plasma/serum and incubate at RT (rotating to keep the GM mixed) for 10-15min. Spin down the GM, wash as per the kits protocol, and then elute your DNA.

The only real problem with the above method is that the DNA which if found free in the serum/plasma tends to be highly fragmented.

Bryan
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Old 10-15-2009, 11:29 AM
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Default Re: DNA extraction Method from Blood

hi
You can take whole blood sample and spin it down at maximum speed so that serum gets separted then use Cell lysis buffer so that cells gets lysed so what remains is leucocytes with free DNA in supernatnat. Then again give a second wash to supernatant so that you get DNA free of any Heme since it may hinder in PCR/RE if you are going for that or otherwise also you need a pure prep. Since you have DNA in eppendorf go for alchol precipitation. Run on gel you get a sufficienct DNA. I hope this would solve your problem . If at all you find any problem please feel to contact.
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Old 10-17-2009, 08:24 AM
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Default Re: DNA extraction Method from Blood

Thank you for this reply, Actually I want to do GWA study from these sample i don't want fragmented DNA, The problem is the Blood is already collected form 60,000 patients from various part of the county. Other thing I have plan to use Qiagen Autopure with robotic arm... but its any pre-treatment to get high pure DNA
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Old 10-26-2009, 04:34 PM
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Default Re: DNA extraction Method from Blood

In this case, you're best bet is a prep from the blood cells. Simply isolate as per any other cell type - you'll get better yields if you can remove the RBCs first. There are a lot of ways to do this - buffy coat separation, dextran sedimentation, etc, but by removing RBC's you'll really up you DNA:cell debris ratio, giving you a better isolation.

Since you're doing GWA studies, I'd avoid plasma/serum DNA like the plague

Bryan
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