I need to extract RNA from cannabis. My buffer extraction is Biozol but when I use nanodrop to identify the concentration and quality of my samples, 260/230 ratio was so low. I think my samples have phenol contamination and my used protocol cant remove it. Do you agree with me? which protocol do you advise to me? did you have any exprience about it?
1) don't be greedy and don't try to take ALL of the aqueous phase after first spin (unless you are using phaselock tubes, which are awesome but are expensive and unless your material is truly precious, not routinely needed).
2) do a second wash with 75% ethanol and break up the pellet with the tip during bth washes.
Some RNA isolation methods use a special lysis buffer with high concentrations of guanidine hydrochloride, instead of high guanidine isothiocyante. This kind of buffer can improve RNA extraction from samples rich in metabolites (such as Cannabis floral tissues).
If it is the phenol that is causing your issues, try removing it using chloroform. Add an equal volume of chloroform to your tubes containing RNA. Quickly vortex and then centrifuge at max speed for 5 minutes. Phenol is more soluble in chloroform than it is in an aqueous solution, so you will be able to extract the phenol using this method.
Also, I have read that RNA samples extracted from plants have a tendency to get contaminated with sugars, since they will dissolve in the aqueous phase with the RNA. This will increase absorbance at 230 nm and reduce your 260/230 ratios. Norgen BioTek has a plant-specific RNA isolation kit that you may want to try out. Alternatively, you could try removing sugars using CTAB.
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