Hello everyone. I'm from Brazil and recently, as you must know, we have no money to buy the most famous dna extraction kits like powersoil and i got to get my goal. So i've already try one protocol using CTAB-NaCl-SDS and PCI (25: 24: 1) but it did not work because there were no bands revealed in agarose gel. Actually, i've tested 2 kits (dneasy power soil and dneasy power soil Pro) using samples that was provided by the company and guess what? did not work. What other alternative should i use to get my total DNA?
Are your samples expected to be low in biomass? Did you try to measure your DNA concentration post extraction with Qubit or nanodrop? Just because you can not see bands when you run your un-amplified DNA on a gel that does not mean that you do not have DNA. What do you plan to do with your DNA and how much do you need? It also depends on the amount of sediment you extracted from, the nice thing about the CTAB-PCI method (or similar) is that you can scale them up to large amounts of starting material (10+ grams) which is usually not possible with kits.
Thank you Utsa Roy for sharing your protocol! I really didn't know about it would be possible to use such a large sediment sample value. I used 0.5g of sediment for the process. And sediment is a kind of diluted sample. Maybe that was the problem. I'm gonna try.
Daniel Lipus I need the total DNA to study diversity of filamentous fungi present in sediments. Sediment is complicated kind of sample because it depends on soil granulometric composition and the river flow and other features. To concentrate the DNA is the difficulty. So yes, I've measured the DNA and it was not so good. As I said to Utsa Roy, I did not know I could use 10g or more and I'm gonna try his protocol. In anyway thanks for helping me.
Hi, Viviane Lopes. Did the Utsa's protocol work for you? Thank you.
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