Hi everybody,
Im trying to isolate RNA from umbilical cord total tissue by nitrogen freezing and powdering and then Qiazol/Choloroform extraction. I have obtained good results with placental total tissue but with cord the problem is that the pellet is very viscous, and when I try to solubilize it in water to quantify, I cannot even to pippete because of this.
Could anyone show me a protocol to do this?
Thanks
Did you ever try cutting a piece of the pipette's tip to increase the volume to be catched at the same time?
I mean, when we do experiments that we must mix tween 20, for example in a buffer, to improve it we do the same, just cut a piece of the pipette's tip.
Good morning Pedro, thanks
The main problem is that I dont think I have really pure RNA, there is something more (hope not DNA but i dont know) that is making my sample viscous. Even more, the ratio 260/280 is so low... about 1.2-1.3. So there is something more in my sample.
But anyway thank you
Why are you using Qiazol/Choloroform some of the RNA isolation kits are better optimised for extracting RNA from tissue. It might be worth using DNase digestion steps to reduce the viscosity.
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