2 replies to this topic

[sleepingbear]

Hi,


I've heard thats adding a proteinase K digestion step during RNA extraction can improve yield. Has anyone had experience using Proteinase K digstion step? Will it actuallyimprove yields? and what concentration do you use the Pro K? The sigma gen elute Total RNA minipep kit suggests 10 ul in 60ul nuclease free water however they don't mention a concentration.

-thanks in advance

[Curtis]

I have recently bought a Miniprep kit from Qiagen. that one also doesn't mention how much proteinase K you need to add!

I think if your sample is cell or virus you wouldn't need proteinase K....perhaps it is only used for fiber-rich tissue

[bob1]

You shouldn't need to add prot K, it is just adding more protein to the solution, which you will then have to remove. If you have a tough tissue such as muscle, I would use a homogenizer such as Polytron to disrupt the fibers.