RNA extraction - Zymogen kit - (May/26/2015 )
I am using Zymogen RNA extraction kit for extracting RNA from brain samples. I have the following questions regarding RNA extraction:
1) I am using a pestle to crush the tissue and it is recommended by the manufacturer to use the Lysis buffer while i crush the tissue. But while using the lysis buffer for crushing the tissue there is foam generation and its difficult to transfer the mixture to a new tube. Any suggestion what can be used alternatively instead of the lysis buffer while crushing the tissue and to avoid foaming?
2) There is a DNAse treatment included in the protocol which i did and i again did another DNAse treatment. Will that harm my sample in any ways?
Lysis buffer helps to stop degradation of RNA during disruption of the tissue. brain tissues are very soft, you can directly homogenize it in the lysis buffer in a tube, or you can grind in the liquid nitrogen, quickly transferring to the tube without allowing the ground tissue to thaw and homogenize with lysis buffer. foam is due to type of tissue, most of the RNA will be in the liquid phase.