Liver is a highly metabolic organ hence you should get enough RNA quantity. If you are facing difficulty in getting RNA then follow this suggestions:

1. Since RNA is very unstable, use fresh tissue for RNA isolation on the same day of sacrifice. If your lab facilities does not allow this, then initially store the tissue in RNA stabilizing solution at fridge for 24hr and then in deep freezer.

2. Use new trizol to crush the tissue, do proper homogenization and during homogenization process frequently keep the homogenate on the ice so that temperature maintains.

3. Use fresh chloroform for differential centrifugation.

4. During isopropanol wash, if you are not getting white RNA pellets after centrifugation, repeat the step by adding more isopropanol and allow to chelate at -20 for 15-20 min before centrifugation.

5. Make sure the life of DNase is not expired.

For detailed protocol you may refer my paper: https://doi.org/10.1016/j.phyplu.2024.100679

Hafeezunnisa Mohammed

Thank you Arghya, I appreciate it.

If you are not getting pure RNA or getting DNA with RNA so you should follow these steps:

• Check your sample collection method freeze the sample immediately to preserve the RNA .
• Use RNA later (sigma) to store the tissues.
• Do all the steps on ice or coolant.
• If you are getting protein contamination so you can try with phenol-chloroform method for RNA extraction doi:10.1016/j.mex.2018.05.011