I am looking for a good RNA isolation method. Most methods I have found take the whole tissue section to make RNA. However, I would like to take a small region of interest from a tissue section to make RNA.
I first dissect out my tissue, then freeze using isopropanol and dry ice. I embed in OCT, then store in -80. From there I am able to take 100 um sections to find specific sections of interest. But I need help dissecting/isolating a small portion of this section for consistent and good quality RNA extraction.
You can simply trim the block to the desired depth, then use a knife to cut the region of interest on the face of the block. If you then cut the section at the desired thickness, the ROI will be separate from the rest of the section.
There is no need to cut deep into the block. Gently scribing a line will suffice to go 100 µm deep.
I will take a dish full of RNAlater or similar solution. Under stereomicroscope and cut the section of tissue as you like. RNAlater protects your RNA. You are going over very long procedure unnecessarily. And also you can store tissue in RNAlater following the instructions with it. Good luck.
Like Jonathan said you can just cut the ROI with a blade, as it seems to be large enough to be visualised by eye (or stereomicroscope). Or else if you have access to a laser-capture microdissection apparatus you can do it that way, once your sections are taken.
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