vortex RNA? - (Jul/14/2005 )
is vortexing RNA bad for the quality of it? I vortex DNA all the time before using it as a template for PCR without any problem. Yesterday I made a new dilution of RNA and used it as a template for an RT-PCR and I only got small aspecific bands (I always get these bands) and only when my input was the highest concentration of RNA. Last time I just diluted everything before purifying RNA so I didn't vortex it afterwards and I got good results. (I got the desired product with an input of "10^6 'till 10^3 and now only aspecific with an input of 10^6-5*10^4).
My guess is that the RNA got damaged by vortexing it.
anyone got any experience on this?
(I know that with RT-PCR a lot of things can go wrong, but I am very positive I did everything else exactly the same as last time).
I always vortex and spin my RNA.. never had any problems there *shrug*