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Homogenizing animal tissue to RT-PCR in real-time


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#1 Lina Salazar

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Posted 16 September 2009 - 09:28 AM

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

Edited by Lina Salazar, 16 September 2009 - 09:30 AM.


#2 Mighty Mouse

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Posted 17 September 2009 - 04:08 PM

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.
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#3 AussieUSA

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Posted 18 September 2009 - 07:11 AM

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,

AussieUSA.




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