Best Total RNA Isolation Kit - For plants, and tiny samples <10mg (May/11/2011 )
What experiences have you had with the RNA isolation kits used for small amounts of plant tissue (<10mg)?
In particular, I would like to know about: RNAqueous®-Micro Kit 50 purifications from Ambion; Mini RNA Isolation Kit™ from Zymo Research; and the MagMax Total RNA Isolation kit from Ambion.
Or, if there are any other kits that have been successfully utilized to isolate total RNA from <10mg of tissue, I would like to hear about those.
I don't have experience working with those exact kits or a low weight of tissue but I have worked with kits from Ambion and Zymo. Which you choose depends some on what plants and tissue you are using and the desired application. I didn't do an exhaustive comparison; in my opinion, Ambion kits give a higher quality RNA than other kits, but the purity is not as good, especially 260/230 absorbance (carbohydrate). The Ambion kit may involve more tinkering to get a higher quality end product with acceptable purity; other kits yield RNA that is suitable for most applications, but if you want to be more sure you have full length RNA for making libraries and so forth, Ambion may be a better choice.
Qiagen, Zymo, and similar plant kits use an additional column that filters out most of the cell wall material; Ambion kits don't use this kind of pre-filter. What I like about Zymo Research is that you can easily buy all components seperately, so you could start out with the plant kit and finish with columns suitable for smaller elution volumes (6ul). You could also just buy their filter column and use it with the Ambion kit.
We've worked with RNeasy from QIAgen and the SV Total RNA Isolation System (Promega). Much better results with RNeasy (higher yield and purity). I agree with David C H that you profit from using a kit with an additional column (for gDNA elimination) especially if you are dealing with a low amount of starting material.
David C H on Thu May 12 14:10:06 2011 said:
What I like about Zymo Research is that you can easily buy all components seperately, so you could start out with the plant kit and finish with columns suitable for smaller elution volumes (6ul). You could also just buy their filter column and use it with the Ambion kit.
From what I understand, a lot of researchers have switched to Zymo over Qiagen because they are similar kits yet more cost effective.