Overdried pellets - RNA extraction - How to recover that jelly? (Apr/09/2008 )

It happens in RNA extraction that we overdry the pellet and it's extremely impossible to resuspend. Does anyone know a quick and dirty way to dissolve it and rescue the experiment? Heating maybe, or is it safe to keep them in the fridge for an hour?

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Hi!
DISCLAIMER: I'm used to extract RNA with kits, so I'm not sure about the characteristics of the pellet we are talking about.
Anyway, it doesn't seem a good idea to me to expose any mixture containing RNA to heat...
Did you try with thermomixer? We have one of those very useful things in lab, so if I was in you shoes, probabily I would try to put the samples for a while at 5-10°C with very gentle shaking (the lowest available), then I'll check if something resuspended...

What do you think?

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heating RNA is a big no no! Try to resuspend it using the normal extraction kit and use the buffer to do so. I dont recommend vigorous shaking. It might disrupt the RNA.

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I don't use kits, because Trizol is the only way (well, only cheap way) to recover some small rnas I need.
Anyway, it's too late (scratches head).... I played with it a bit. I left it in 55 Celcius for 5 min, it worked and I still had high yield. I saw a procedure like this somewhere in the net. Hopefully subsequent reactions will work OK.

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Hi Telomerase, I always incubate my resuspended RNA pellet at 60C for 5min after Trizol extraction, and have always had good yields and good qualitiy, so your RNA should be ok for downstream applications. You can always run a gel to confirm.

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I used to follow a quite established protocol to extract my RNA. The final step after i precipitate, I dissolve my RNA with RNAse free H2O and heat it (but not shaking) for 10 min at 65C.

see this: http://www.path.cam.ac.uk/~toxo/Protocols/...20ISOLATION.pdf

It works fine. From agarose gel, Nanodrop and Bioanalyzer result, my RNA quality is always very good.

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