I would like to store adult small strongyle specimens until they can be morphologically identified. In addition, i would like to return to identified specimens and attempt to sequence some genomic dna. Any thoughts on feasibility? Any advice would be much appreciated.
80% alcohol (ethyl alcohol) is a better option.
This paper could also be of help (Structural Restoration of Nematodes and Acanthocephalans Fixed in High Percentage Alcohol Using Dess Solution and Rehydration by Naem, Pagan & Nadler in Journal of Parasitology (2010) 96(4) ).
If you've got -80 freezer space snap freezing might be your best option, otherwise I'd probably go for ethanol - reasonable tissue preservation for gross ID of parasites and your DNA will be ok for downstream PCR or NGS applications. Don't put them in formalin if you want to recover DNA (absolute nightmare trying to get DNA back out). Good luck!
Thanks, all for your answers. We are currently stuck preserving nematodes in formalin. If they are stored in ethanol they shrivel up and cannot be identified even after trying to rehydrate them. Haven't tried snap freezing yet. Any other ideas?
Glycerol? or RNA later? both good for DNA recovery but I guess you'd have to try and see what they looked like afterwards. In the past we've also ended up splitting samples - part in formalin and part in something else for the DNA. It is possible to get DNA out of something that's been in formalin but I have never had a lot of luck with it
Hi- I work with large marine mammal parasitic nematodes for DNA work and have found the best storage conditions to be -80C in 1x phosphate buffered saline. However I have been able to recover DNA from specimens stored at room temperature in ethanol or formalin. second to pbs I would suggest 70-100% ethanol at -20.
Hi. I have been successful in studying morphology & extraction of DNA & PCR of Insect nematodes by storing it in 10% Glycerol at 4C. Not sure for other nematodes but you can try & check.
Hi Caroline,
I have extracted DNA from a dog parasitic nematode, which is stored in 70-90% ethanol at room temperature or fridge. I used QIAgen Blood & Tissue Kit. The DNA sometime was recovered but most time was not recovered. That means my samples' DNA was degraded before I extract when they were stored in room temperature or fridge?
Thanks
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