Standard protocol for DNA purification from Frozen Blood??

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Standard protocol for DNA purification from Frozen Blood??

Postby ashsu89 » Mar 04 2012 3:48 am

Does anyone have a reliable standard DNA purification protocol for extracting DNA from frozen blood samples? I have limited experience in bio techniques and have never done this type of procedures before. I was wondering if anyone have good suggestions of simple and reliable procedures I can follow without the use of phenol? I currently have frozen blood samples that are stored at around -80 degrees in Quiagene PreAnalytix Blood DNA tubes. However, since I could not purchase the kit, I need a standardized procedure as well as a list of reagents/buffers I need to help me purify the DNA from the blood samples.

Anyone have suggestions about what I should do?? I'm getting pretty helpless and desperate in trying to figure out a way... Thanks so much!!!
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Re: Standard protocol for DNA purification from Frozen Blood

Postby biochembug » Mar 17 2012 8:49 am

Hi

What for you require the DNA? Regular PCR?

You may find several protocols which are based on proteinase K digestion followed by phenol extraction. For removal of proteins, you will have to do a phenol chloroform. I guess if you do not want column you will need a phenol extraction. I have also seen some papers showing direct PCR from blood samples. Google it or visit
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC332355/,
http://ps.fass.org/content/86/1/102.full,
http://www.clinchem.org/content/47/9/1701.full.

Hope this helps!

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