How can I improve low 260/230 ratio in RNA extraction with trizol??

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Mark Ian Fowler

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).