I have been using Biorad RNA synthesis kits which gives better RNA yield and quality from most of the tissue of breams, silver pomfret, seabass.Muscle tissue RNA yields very low and the quality many a times falls below 1.8 (260/280).
What best modification one can suggest?
Thanks in anticipation.
azad
Dr. Azad, Use trizol, it will give you a better yield…you can also scale it up, if you have more tissue
As the other respondent stated, Trizol (also sold under other brand names) is a highly performant method for extracting RNA from a variety of materials including muscle.
Personally, I would recommend RNAzol RT. It provides very high RNA yields with a simpler protocol. I have used it to extract RNA from mouse skeletal muscle for reverse-transcription gene expression experiments. I was able to get reliable RT-qPCR results even for low-copy-number transcripts.
I have isolated RNA from Antarctic fish liver tissue, with a very low tissue amount ranging from 50 to 100 mg. I used Trizol reagent and got good yields of RNA with high quality, suitable for downstream applications like real time PCR. Please ensure the tissues are stored in -80 freezer and need to be snap frozen in liquid Nitrogen after harvest and before storage in deep freezer. Use liquid nitrogen to grind the tissue using pestle and mortar. I have used tissues stored for over 6 years and obtained good results.
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