i have extracted RNA from wheat shoots using trizol method.
ran extracted RNA on agarose gel, i am confused about the bands.
help would be appretiated..
First off, you can read off the molecular weights of your bands using the molecular weight standard markers on both extreme sides of the gel. It is highly desirable to run the molecular weight standard markers on BOTH the leftmost and rightmost lanes so you can use a ruler or any straightedge to
connect identical standard marker bands, and this makes the reading of your own bands much easier and more accurate than if you just have the standard marker bands on one side.
Did you denature the RNA before loading (or use a denaturing gel)?
I have essentially no experience of plants, but I am guessing that the chloroplast ribosomes (being more prokaryotic in sequence) are 16/23 and the cytoplasmic ribosomes are 18/26, so seeing four prominent bands is what you'd hope for.
If you want to improve resolution, you can denature RNA by mixing with formamide (1ul RNA, 2ul 6x sample buffer, 9ul formamide), heating to 65 degrees for 5mins and then crash cooling on ice. This RNA can be loaded on a standard agarose gel, saving you the effort and hassle of making a denaturing gel.
Hi dear Hafiz Imran Fakhar
you must keep that in your mind that when you load your RNA sample on electrophorus agarose there are 3 RNA band available 28s,18s and 5.8 and at the top of your screenshot there are DNA band and at the next step you must use DNase to eliminate DNA
an also I think you have a bad RNA extraction you need work on it
good luck
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