This is a time study that I am performing where the cells are exposed to TNBS in SFM at different time points. I'm then waiting 24 hours in normal media and then harvesting the cells for DNA and RNA. When I'm doing the RNA, I added 1 mL of Trizol straight into the wells and let set for 3 min. Pipette up and down and then another 5 min hold. Moved the 1 mL of trizol and cells to a 1.5 mL tube and added 200 mL of chloroform. 10 min at R.T. and centrifuge at 10,000 for phase separation. Removed the upper phase to a new tube and then added the 500 uL of isopropyl and went another 10 min before centrifuge 14,000 for 15 min. Then wash and dry the pellet. This procedure is not working that well for me. Does anyone have any suggestions of where I can possibly improve or optimize this?
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA