Acid phenol - (Jul/07/2007 )
did any one know how to made acid phenol
- could I saturate it with water
- or with sodium citrate ph 4.5
and what's better for guanidine solution (TRI reagent)
use the sodium citrate, you need a buffer to main the pH.
A better solution then trizol, well you could add a third chaotropic agent like urea or KI. Maybe specialise the solution for the cell type you are using, adding CTAB or something similar for plant extraction, could improve performance.
However, I think some improvements can be made by changing the protocol. I got better DNA (I use the sister product DNAzol) by changing the protocol, prombe cells produce far more debrie then mammalian cells.
I use it for recalcitrant plant tissu
what kind of buffer could i use
citrate buffer is fine.
but for recalcitrant plant tissue, well I can't really help as it is not my area of experties. You will probably need more then Trizol. I would suggest using liquid nitrogen to freeze the tissue and use a mortar to break it down. As for the polyphenols and such, well you need to play around with CTAB (maybe DTAB) and PVP. The web has many protocols for RNA extraction from plants, I you suggest you go look at them for inspiration