RNA extraction from flowers - (Sep/03/2007 )

Hi! Have anyone successfully extracted RNA from flower (esp: orchids). I have tried so many times, but obtanied RNA sample contains so much of pigments (no clear RNA obtained). Can anyone help me!


Wijen
University of Malaya

I used kits to extract RNA from vitis leaves, which are considered a quite difficult matrix.
I had pretty good results.
What method did you apply?

The Chang method for pine works quite well for most tissues rich in secondary metabolites (Chang et al, 1993, A simple and efficient method for isolating RNA from pine trees, plant molecular biology reporter 11, 113-116). I can't find a pdf, unfortunately.
This method was suggested to me some years ago in a forum similar to this (was called 'the rna forum') and sent in shortened word version. I attach here this version, hope it can help.

Hi! Just to add in another query here. I am trying to extract RNA from Polygonum minus root (a type of local herb), not from flower. My target plant is kind of rich with secondary metabolites also. After failing to get any RNA extracted by using QIAGEN RNeasy Plant mini kit, I have shifted to conventional methods. I had tried out with several methods, which is describe by (1) Irene Pateraki and Angelos K. Kanellis 2003, (2) Abbas Alemzadeh et al. 2005, (3) a modified phenol-SDS method. I also tried using Trizol reagent but no RNA yields at all. So far, I only get very faint bands by using the method described by Irene and Angelos. However, recently, I couldn't get any RNA bands although I used this method. I change all my solutions, assuming that some of it may contaminated with exsogenous Rnase but still this couldn't help. May I know any other kits or methods which is suitable for my plant? I will try the method describe by Chang as well.

Another question is, will the type of plant culture medium affect the content of RNA or the RNA isolation process? Because before this I used to culture my plantlet in solid medium but since it took about two months to grow, so I had changed to liquid medium in order to let the plantlet to grow faster. I noticed some changes in the plant morphology also. However, once I changed to liquid medium, I couldn't get any RNA at all. Please help! Many thanks in advance!