mistakenly use RNeasy spin column while extracting RNA - (Oct/15/2008 )
I made a terrible mistake today, when i extracted RNA from white blood cell using RNeasy mini kit, i should use QIAshredder column to homogenize cells, but i used RNeasy spin column mistakenly, and continued all steps only with RNeasy column including on-column DNase digestion.
Does anyone know what possible consequences would I got? low RNA yield? contamination?
I measured the RNA quality by NANO-DROP ND-1000 spetrophotometer, the resluts as shown below
Sample 1: Concentration 444.3 ng/ul, A260/A280 ratio 2.01
Sample 2: Concentration 125.5 ng/ul, A260/A280 ratio 2.19
Should I trust these results?
Thanks a lot!!!!
Given that your RNA isolation has decent concentration, there is nothing to worry about. I guees when you mistakenly used the RNeasy column for the shredder, and did the centrifugation, the RNA did not bind much to the column because the condition did not favor the binding (before adding 70% ethanol). I think the only consequence is that you lost some RNA. That is it.
Thank you for the explanation!