Does anyone know an improved protocol to extract RNA from A. thaliana leaves?
Thank you!
Dear Sir. Concerning your issue about the suitable protocol for RNA extraction ( from A. thaliana leaves). Arabidopsis thaliana is widely used in the research of plant molecular biology, but RNA isolation from certain tissues is difficult. For example, siliques and dry seeds require complex and time-consuming methods due to the high contents of polysaccharides and secondary metabolites (1–3). One way to treat such difficult tissues is to use a high-salt extraction buffer, an example being LiCl, which was used in previous studies (1,2). Here we report that the use of a high-sodium extraction buffer is more effective in RNA isolation from these tissues. Our method consists of a two-step extraction with organic solvents, isopropanol precipitation, and LiCl precipitation, if necessary, after sample extraction and is simple enough to be routinely applied to other tissues. I think the following below links may help you in your analysis:
http://www.biotechniques.com/multimedia/archive/00037/BTN_A_04374BM03_O_37019a.pdf
https://www.genetargetsolutions.com.au/wp-content/uploads/2015/02/QuickGene-Application-Data-Total-RNA-Extraction-from-Plant-Tissue.pdf
http://file.scirp.org/pdf/AJPS_2014101016181867.pdf
Thanks
Try this kit method to get good results
http://www.sigmaaldrich.com/life-science/molecular-biology/dna-and-rna-purification/plant-genomic-dna.html
The kit based method is working nicely , or even by trizol method ,you can get good amount of RNA from leaves of Arabidopsis thaliana.
you can use trireagent or rnazol, you can get good amount of RNA, if you have some problems with A(260/280), sodium acetate metodology is the best.
Good luck.
You have to order the kits for RNA extraction, that is very important specially if you are going to work with cDNA
Thank you all. I have done RNA extraction with TRIzol reagent and it worked really well. Very rapid method also, the quality and amount were great!
So, finally, what is your extraction protocol? Could you please describe it in detail? Thank you in advance!
Anh H. Ngo, thank you for your suggestion. I will try with the TRIzol reagent!
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