I am trying to extract RNA from leaf of White Calla lily by using Trizol method. By using this method, i am getting good concentration value and 260/280 ratio on nano drop measurement as well, however the value of 260/230 is very low. Furthermore, i am not getting clear bands of RNA on gel electrophorosis as well. So, if anyone is using Trizol methods to extract RNA from White calla lily, i would like to get some suggestion to get better quality of RNA.
In my experience, the key to getting good RNA out of plants is all in the sample prep. I would flash freeze the tissue in liquid nitrogen & grind it to a fine powder in liquid nitrogen. Once frozen, you can store the tissue or powder in the -80. I had great success extracting RNA using a Qiagen plant RNA kit. Different method, but it works really well.
I have been using the Sigma Tri reagent to extract RNA from some ornamental plants, following the instructions in the manual. I don't know if this is the one you are using, I really enjoyed working with this one. But is necessary to standardize some things in its methodology, for example: temperature of centrifugation (I use 4ºC), allowed time of samples to stand in room temperature, drying of RNA pellets, elution solution and dissolution of the RNA pellet. I got good results for many samples. For materials that were not successful with this reagent, I used the Total RNA Purification Kit (Norgen Biotek Corp.), and also the CTAB method was efficient for some plants.
Bijay Sharma Subedi Since you do not see clear bands on agarose gel, your RNA preparation is not clean and might have RNase contamination. Always use DEPC treated material for RNA extraction work and also try to have separate work bench for this work.
The next important thing to get the good quality (as well as quantity) of RNA is sample preparation and I fully agree with Katie A Burnette
Follow what she has suggested in this regard. I also prefer using Qiagen RNeasy Plant Mini kit for isolation of RNA from plant tissues.
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