I have tried to extract RNA from fish semen and testis using the RNEasy kit and also have the PureLink RNA kit. I have tried a few modifications I've found online, but the RNA comes either highly degraded, despite what a simple gel run would suggest, or contaminated with other biomolecules. When trying to initially homogenize the tissue it becomes extremely gelatinous and therefore impossible to pipette, and unfortunately I do not have sophisticated equipment to use beads and other methods, although I am inclined to think the sample would still be too gelatinous for this method. I haven't found many references using liquid nitrogen to homogenize the tissues, so any past experiences would be highly appreciated. Also, if anyone has some suggestions for any of the two kits' protocols it would be appreciated too. I've read the addition of AGPC could be useful in this case.
Thanks to everyone in advance!
I'm not sure if this helps, but there is a product line called DNA/RNA Shield by Zymo Research which will lyse cells and protect the nucleic acids for downstream use. It's compatible with normal RNA extraction kits. We harvested mammalian cells with it and were able to extract the RNA out of the solution for qPCR. There are guidelines for how much to use for the number of cells you need to lyse so it may help with your viscosity problem.
Did you try to load your sample on a shredder column before the extraction?
it helps with saliva.
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