Hi Aradhya Tripathi,

We have some methods to isolate RNA. However, Trizol method is very efficient. I have used Trizol for many plant and I have good RNA quality. In my experience, you will use less than 100 mg sample for RNA isolation based on manufacturing protocol. Homogenize tissue sample need to be very fine by grinding with liquid nitrogen.

When you do gel running, buffer, gel ... need to be free RNase. RNA degradation happens during isolation process and also gel running.

PVP (polyvinylpyrrolidone) not PVPP could be used for RNA and DNA extraction to remove phenolic compounds

Hi, take a look to this link, you may find it interesting.

https://www.researchgate.net/post/How_can_I_get_rid_of_polyphenols_in_olive_leaves_RNA_extraction

I agree with Phat Tien Do. Trizol woks well. It's very important to frozen and grinder your sample in liquid nitrogen as soon as possible. After adding Trizol, make sure mixing your samples with the Trizol well.

Hi,

Ambion provide a product "plant RNA Isolation Aid" ref AM9690, you can add this react to your lysis buffer!

Goog luck

Best regards

pierre

Hi I had tried different methods eg. CTAB, Trizol and commercial kits, we found that Spectrum Plant Total RNA kit from Sigma gave a good quality RNA and this kit also work well with any hard handle tissue, and of cause the key step is the way you grind the tissue, Yingzhong gave a good suggestion in handle and grind the tissue.  Give it a try! 

I have been using the RNeasy Plant Mini Kit from Qiagen for many years with good results. However, for carbohydrate-rich tissue , the 56C incubation step should be omitted. We flash-freeze our tissue in liquid nitrogen and then grind it to a fine powder with mortar and pestle.

Depending on the species and physiological state, some chaotropic agents like Trizol and guanidinium causes complexes with secondary metabolites that interferes with RNA solubility.  There are some detergent based extraction methods that resolve this problem.  On old but solid procedure can be found in Hughes, DW, and G. Galau, 1988.  Preparation of RNA from cotton leaves and pollen.  Plant Molecular Biology Reporter 6:253-257.

I have successfully used RNase Plant Mini Kit (Cat. No. 74903) by Qaigen with duckweed (Spirodela polyrhiza).  They include two types of extraction buffers in their kit in case secondary metabolites are an issue with your plant species.  You really need to try both types of extraction buffers to determine which one is best for your plant species.

Article RNA and DNA isolation from recalcitrant plant tissues

Article Preparation of RNA from cotton leaves and pollen

You can take fresh tissue samples and crush them at the same moment without storing. If you are collecting them from faraway places try not to dehydrate m before crushing them. After crushing them, directly introduce them in a vial containing a liquid (Trizol or lysis buffer) according to your protocol. This helps to protect RNA from denaturing as the solution will bind it once it comes in contact with the RNA. And you are looking for PVP40. 

Quiagen RNeasy plant mini kit seems to be ideal from my experience