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RNA extraction/trizol

rna rnaseq

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#1 Tara Burns

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Posted 21 August 2015 - 05:05 AM

Hello all,

 

I am trying to extract RNA from embryonic mouse hearts.  I have a few questions, what are people's methods of disruption/homogenization-we don't have any tools for that right now, I am wondering if there is anything cheap I can buy.  Also, while I do the dissections, is it possible to put the tissue immediately in the RLT that comes with the Qiagen kit?  I tried putting it in RNA later, but when I tried to homogenize with a tube mortar and pestle, I couldn't grind the tissue because it got so hard in the RNA later solution.  And finally, do people find that the trizol method is better than the RNA kits from qiagen, and if so does anyone have a protocol?

 

Tara



#2 mdfenko

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Posted 21 August 2015 - 05:40 AM

heart muscle is notoriously tough. it is very difficult to homogenize with mortar and pestle.

 

possible remedies are:

 

freeze with liquid nitrogen, powder the tissue with mortar and pestle or tissue pulverizer then suspend in an extraction buffer and homogenize as you tried before

 

or (if you have the funds) obtain a generator probe type tissue homogenizer like this: omni international general tissue homogenizer


talent does what it can
genius does what it must
i do what i get paid to do





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