I am about to take blood samples from my patients, extract mt-DNA from those blood samples and then run PCR. My question is how to preserve these samples?
Okay dear thanks, I need to know who would we extract mt-DNA from it, and when I would take blood from the patients then how I can preserve it as the minute I would take blood I would not go for extraction directly it will take a few weeks then I will extract so what should I do in this regards should I freeze the blood or what?
Hi Aamir, we use HISTOPAQUE-1077 to obtain polymorphonuclear cells from peripheral blood (lymphocytes and monocytes). We use either EDTA or heparin tubes to obtain blood and then process the blood within two hours (four hours max). Once the PMN's have been obtained (this process takes around 2 hours), we harvest the cells in Tri-reagent and freeze at -80 until we are ready to process the sample.
From my experience, I would not recommend freezing whole blood before isolating leucocytes. There may be other methods but I am not familiar with them and, can therefore, not recommend them. This paper may be useful however http://www.clinchem.org/content/47/9/1701.full
I hope this helps.
You can preserve the blood samples at room temperature for two weeks if you follow this protocol:
http://www.preanalytix.com/product-catalog/blood/dna/
You need a biosafety level III facility to work with human blood samples because you never know what kind of infectious disease (hepatitis, HIV etc.) it may carry. Besides, if you want to publish your results in a reputed international journal you need to collect the samples lawfully and with proper consent of the patients. Moreover, your project needs to be approved by a recognised Research Ethics Committee.
Firstly you go through Histopaque method of seperation and then you make pellet at higher speed centrifugution that is necessary for pelleting mit DNA then you separate this organelles and applied process as attached paper
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