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Homogenizing animal tissue to RT-PCR in real-time - (Sep/16/2009 )

How to homogenize an animal tissue (lung human) in order to make a RT-PCR in real time?

-Lina Salazar-

We use mouse brain tissue in our lab. Following dissection we place the tissue in RNA Later. Then it's homogenized using a Wheaton Dounce Homogenizer in Trizol and then use the RNeasy system from Qiagen.

-Mighty Mouse-

Similar to Mighty Mouse, we add the tissue to RNAlater, but quick-freezing tissue is fine. Homogenize either in Trizol or any commercially available RNA prep kit (we use Qiagen RNeasy). I might recommend DNase-treating prior to preparing cDNA as tissue is more "messy" than cell line. And treat as normal after that.

Hope this helps,