Hi researchers
i plan to do a study of genetic polymorphism associating with primaquine therapy in our population. I was wondering how much of venous blood should i take from patients for DNA extraction before storage in consideration of keeping a few as a back up. And how does the extraction go? Extract the whole of volume and transfer them into few aliquots. I plan to do 3x at a time from 3 aliquots and find the SD for one patient and i should have remaining 3 aliquots as back up for future use, just in case.
Thanks
Liza
Hi Norliza
I used salting out method for DNA extraction, and after that I stored them in -20. Their quality is good enough for PCR. I used this protocol but with a little bit modification.
- Rousseau F, Rehel R, Rouillard P, DeGranpre P, Khandjian EW. High throughput and economical mutation detection and RFLP analysis using a minimethod for DNA preparation from whole blood and acrylamide gel electrophoresis. Hum Mutat 1994; 4: 51-54. –
Good luck
thanks all for the advice. Now if i plan to take blood samples at hospital which is far from lab, then what is the best way to keep them in regards with logistic aspects and ensure the DNA is still intact and in a good quality? i may assume that i could possibly keep sample in my own house.
Use EDTA as an anticoagulant and then save the samples at -20C. it is better to extract DNA as soon as possible.
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