5 replies to this topic

[BottegaVeneta]

Hi,

I've been having trouble getting clean RNA with Trizol.  I try to do everything carefully (ie: not touch the interphase after chloroform extraction, etc).  I also resuspend my final RNA in water, but I heard you can resuspend RNA in TE buffer and get better A260/280 readings.  Is using a column better than the more time-consuming Trizol method?  

Thanks!

[fishdoc]

BottegaVeneta, on Oct 7 2009, 02:55 PM, said:

Hi,

I've been having trouble getting clean RNA with Trizol.  I try to do everything carefully (ie: not touch the interphase after chloroform extraction, etc).  I also resuspend my final RNA in water, but I heard you can resuspend RNA in TE buffer and get better A260/280 readings.  Is using a column better than the more time-consuming Trizol method?  

Thanks!

I use the RNeasy kit for bacterial RNA. It works great. I had tried using Trizol a few times, but it never worked all that well. The RNeasy kit takes 30-60 mins and always gives a good yield.

[BottegaVeneta]

I wish there was a cheaper alternative that's just as quick and easy as a kit.  I have to do RNA time courses, and the price really adds up.  

[littleaxt]

Hi!

I always had problems with gDNA contaminants when using the colums, so I changed to Qiazol (more or less the same as Trizol) and got much better reults. but in the end I apply both no, Extraction with Qiazol and after DNase I digestion I make a clean-up with RNeasy columns. I get really pure, high quality RNA but it is also very expensive :-(

[Johannes]

in case of miRNA a lot of recent papers describe better yield using TRIzol than any available kit.
we are using TRIzol for RNA extraction and qRT-PCR on miRNAS, too. No Problems so far.

[moerae]

We use both Trizol and columns in our lab.... Kinda an overkill but RNA is really clean afterwards.