I have problem with RNA extraction from soil. My RNA have low yield and purity so i want to give a proteinaseK treatment to the reaction. How much i should add proteinaseK and how to use it? Thank you
A typical working concentration for Proteinase K is 50–100µg/ml.
Proteinase K is used during DNA extraction to digest many contaminating proteins present. It also degrades nucleases that may be present in DNA extraction and protects thee nucleic acid from nuclease attack. Can be added to extraction procedure to optimize RNA yields. For isolating cytoplasmic RNA , centrifuge the cell lysate, remove the supernatant and add 200ug/ml proteinase K and SDS to 2% (w/w) then incubate for 30 minutes at 37oC. For more details consult https://agscientific .com --- 2019/04
If you are using a ready made kit, there should be manufacturer’s instructions on how much proteinase K to add. Even if you are not using a kit, these manuals can provide a helpful guide on the proper ratio of reagents to add for your extraction.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA