My lab is working in a project where we collect residual material from thyroid FNA biopsies.
We collect the residual material by "washing" the syringe in RNA later solution (in the eppendorf) right after the FNA biopsy is done.
Since it's not a lot of sample to start with, considering that the material is diluted in ~200 uL of RNA later, how could we improve the concentration yield for DNA and RNA?
We've used Invitrogen PureLink and Qiagen QiAMP extraction kits for DNA isolation, but our concentrations are around 1-2 µg/ml most of the time and OD varies a lot.
As of RNA extraction, we've tried TRIzol + Chloroform protocol, but same concentration problem happened.
Has anyone worked with thyroid FNA biopsies sample before? Is this a normal concentration? Any tips on improving them?
Thanks!
How did you do sample disruption and homogenization before RNA extraction?
I know that the extraction kits have the ability to lyse the cultured or certain tissue cells, but in many cases, the RNA extraction from tough tissue cells still requires a robust cell disruption process.
You may want to try TissueLyser using mechanical shaking which provides complete homogenization before any kit or TRizol extraction.
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