I am trying to extract RNA from stem cells encapsulate in the hydrogel, but I have a problem getting reasonable yield numbers. Does anyone have any idea why? Would be much appreciated.
Few adaptations may be recquired from protocols often designedfor 2D growing cells. What is your hydrogels made from ? How many cells do you expect to recover ? what are your yields and 260/280 ratio ?
Dear Anoosha,
I normaly extract RNA from PEG based Hydrogel, and what we normally do is to freeze the biomaterial with engulfed cells in N2 liquid and to crumble in melting pot in order to obtain a very fine powder and than we proceed with classic trizol protocol or kit for RNA extraction.
I hope this will help you.
depends on what your hydrogel is. I would first attempt to extract directly in TRIzol. If that doesnt work, you may have to digest/dissolve the hydrogel to release the cells.
My hydrogel is Alginate, I'm making hydrogel beads about the 1 mm diameter with 5000 cells per each. I tried dissolving the alginate ang getting the cell pellet out but still I get numbers of 260/280 about 1.3-1.4 which is way below the acceptable range .
Hadi, have u ever gotten PEG contamination? How did you get pure RNA?
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