I'm trying to extract fungal RNA from soil samples, I've used the NTES method, which is supposed to be more effective than the Trizol method, but I had no good results. Can anyone help me with this? I think my problem is at the initial steps. What's a good way to break the mycellia at the beginning and avoid RNA degradation?
Hello Paulina, check if this article helps you...
http://link.springer.com/article/10.1007%2Fs00253-012-4209-7#page-1
Hola Paulina,
RNA from soil is difficult. But do you know the following paper:
Persˇoh, D., Theuerl, S., Buscot, F. and Rambold, G. (2008) Towards a universally adaptable method for quantitative extraction of high-purity nucleic acids from soil. J Micro- biol Methods 75, 19–24.
They describe a very good method providing good RNA.
All the best,
Dominik
Hi Paulina
you could try the MoBio kits:
http://www.mobio.com/soil-rna-isolation/
the main problem with soil isolation is the removal of the many PCR and enzyme inhibitors present in soil. I have no experience of the RNA kit but their DNA kit works well for applications including PCR and restriction enzyme digestion. I think they do free samples if you want to try it out.
best
Paul
Hi! Thanks for your suggestions, I may be trying the kit.
If Dominik Begerow could pass me the article, I will be glad.
Thank you very much.
Hi Paulina
and for sure this kit will work, same was the problem when we started our RNA work, now its moving fine..Kit is awesome...
and we can also make DNA from the same fraction, using just one step of filter dissolution
I know this sounds crazy but try Adolph's meat tenderizer. I had trouble isolating DNA from onion cells and it worked. I don't know if it will harm RNA.
Thank you, I tried today the traditional method again, with a few changes at the beginning, I hope it will work. Then I'll try to convince my PI to buy the kit...
Qiagen Kit also gives good RNA quality. You can preserve the soil in RNA later solution at -80 degrees.
You can use MOBIO soil RNA extration kit, it work fine recently we have used this kit,
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