Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue
You were able to see 18S and 28S rRNA bands which indicates your RNA is mostly intact. In two of your samples, you see a smear in lower molecular weights which might indicate partial degredation in those samples. When isolating RNA from tissues, keep in mind that there are way more RNases in tissues than in cells. Do not use more tisues than needed. Tissue type is also important. For example, spleen has more RNases compared to most of the tissues and it is hard to get great RNA from it.
Your RNA prep looks just fine to me! You expect to see two rRNA bands and a lower weight mRNA smear. Some of your preps had a higher yield than others, but that's normal too. Continue with your experiment :o)
Thank you so much Sibel Cal and Katie,
This RNA extracted from fresh breast tissue, my concern is that the 28s bands are not twice the size of 18 s bands.
I will like to suggest the buffers. I Will suggest the transfer buffer
I've seen a better result by adding 40 mM DTT in the extraction buffer.
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