A simple sodium acetate precipitation of the RNA will clean up your A260/230 ratios. Here is the method I use:
- Add ultra pure water make up RNA samples to 100ul in total
- Add 300ul 100% EtOH & 10ul 3M Sodium Acetate per sample
- Vortex + incubate overnight @ -20oC
- Centrifuge full speed @ 4oC for 30mins
- Pipette off supernatant.
- Add 500ul 70% EtOH + mix
- Centrifuge full speed @ 4oC for 15-20mins
- Pipette off supernatant & air dry for at least 15 mins [check that no EtOH remaining]
- Resuspend in 30ul or 12ul (depending on RNA concentration)
- Enjoy good A260/230 values!
Give it a go, it works really well. Just remember the RNA pellets will be difficult to see (pellet paint can help).
I would do an ethanol precipitation cleanup for your RNA, definitely allow to precipitate at -20 overnight and do at least 2 washes with ethanol prior to air drying. I usually also heat at the end to get better resuspension.
https://cshprotocols.cshlp.org/content/2020/3/pdb.prot101717.full
- Badges
- Science topic
- Similar topics
- Microbiology
- Assays