I have to isolate RNA from Populus nigra roots for real time PCR analysis. I use commercial Kit (Sigma). After some modification RNA isolation from leaves work well, but for the roots the yield is too low and then the real time PCR analysis are not good: high Ct and not good repeatability. So, how can I improve my protocol to obtain higher yield with root tissue?
Hi Muriel,
in our Lab we also tested the coemrcial kit from Sigma to isolate RNA from Quercus suber roots for qPCR analysis and didn´t work out as well. So, we have been using a more time-consuming protocol (Hot-borate method, Wan, C. Y. and Wilkins, T. A., 1994) yet with very good results, not only on the yield but more importantly on the quality of RNA. This protocol has been used for Vitis leaves, Quercus leaves and roots, fungi, flowers, etc. If you don't have any other suggestion, this is a very good method.
Hi Filipa, thank you very much for your advice. I'll keep it in mind.
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