For DNA extraction from cell line or PBMCs, which protocol is suitable for this experiment? In my opinion, commercial kit such as DNA extraction kit with silica column is convenience and providing the high yield and purity of extracted DNA but other people tell me phenol-chloroform extraction is better than silica column kit. In my experience,I prefer to use DNA extraction kit rather than do phenol-chloroform extraction. I would like you to share your experience and discuss in this point.
Actually my opinion is method should be selected according to your requirement. For example if you just need DNA for PCR, commercial kits will be the best as it is fast and pure from contaminants. But if you need more yield such as for a digestion, hybridization you can go to manual method. Here you can adjust the sample amount and reagents volumes as you need. One time I want to extract more DNA for many techniques. So I adjusted my manual method to yield 10 times more yield of DNA from single extraction. You cannot optimize any reagents from kit to make what you want. But using manual method it can be done.
http://www.hindawi.com/journals/bmri/2009/574398/
Review Article
DNA, RNA, and Protein Extraction: The Past and The Present
Journal of Biomedicine and Biotechnology
Volume 2009 (2009), Article ID 574398, 10 pages
http://dx.doi.org/10.1155/2009/574398
Article should be helpful in selecting the method of extraction
Thank you very much Prof. Nilmini Kumari and Prof. Mushtaq Ahmad for your advice.
It really depends on your downstream purposes with the extracted DNA. If it is just for screening purposes using PCR, even boiling method will work. If it is for DNA-sequencing, you might want to use a good conventional kit, like Qiagen DNA extraction kit.
Dear Prof. Jia Xiang See,
My purpose for DNA extraction is to study SNPs by qPCR. I think this protocol needs high yield and purity so DNA extraction kit is the best, isn't it ?
Hi, I'm just a graduate student, not Prof. Yes, in that case, you will need high purity, and many commercial DNA extraction kits can certainly fulfill your purpose.
There is no gold standard in DNA extraction it depends on the objective of your work.for quantity according ti literature, boiling method is preferred but for quality kits is suggested.
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