I am using RNAeasy Plant mini kit (Qiagen) to extract RNA from Arabidopsis seeds. Can anyone suggest which lysis buffer works best with seed RNA extractions? RLC or RLT?
Using kits for RNA extraction from seeds ends up with poor yield and quality, most of the times. Qiagen RNAeasy kit had failed to give me any good RNA from Arabidopsis seeds. Since quite a lot of seeds are invested for RNA extraction, I would recommend using trizol-based conventional methods. I have been using a method given in the following link. It's a bit lengthy, but assures good quality RNA with decent yields.
http://onlinelibrary.wiley.com/doi/10.1002/biot.200900211/abstract
Thank you Yadukrishnan. I agree. I always end up with poor quality RNA. I have tried a few different methods and haven't had success yet. I will check out the link you posted. Thanks again.
Keep in mind that Arabidopsis seeds have a very high oil content, which makes it difficult to extract any nucleic acids.
Check out this online protocol:
https://www.jove.com/video/55008/an-efficient-method-for-isolation-highly-purified-rna-from-seeds-for
RLC is better and used generally to all tissue except those tissue cause solidification during RAM isolation..
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