The Geek in Question
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Denial…or not.
Posted by on February 19, 2010
I think I have finally been smacked across the cheek by the hand of reality.
The scope of my project –
the ridonculous breadth of taxa, the INSANITY of the geographic scale (I mean, THREE ecoclimatic zones…THREE?!?!?!), the human resources/training requirements, travel logistics, equiment needs, AND the challenge of keeping all participating investigators happy with the output (um, anyone have any great ideas for maintaining integrity of arthropod DNA, when said arthropods have been captured in the field and sat in preservative of some kind for 4-14 days? Apparently 80% EtOH and salt water are no-goes…O NOES!) –
all of this amounts to one Geek going AAAAAUUUUGGGGHHHHH over her sampling protocol.
(Followed quickly by “WOOT! REAL SCIENCE, YAAAAAYYYY!!!” And this is definitely the overriding sentiment.)
Hm. Does that make me in denial? Or maybe in denial about my denial.
Bah, either way, I’ll get this all sorted out. By May.
AUUUUGGGGHHH!!!! WOOT!
Sounds like quite the project! Depending on what sort of DNA work you’re looking at doing (genomic vs mitochondrial COI), and the time available to you for working with collected samples, you might be able to use 95% EtOH and exchange the alcohol a couple of times throughout the 4-14 days. I’ve gotten fairly consistent success with genomic genes using 95% EtOH as long as I make sure the alcohol is changed on the return from my trip and the specimens are placed in at least a -20C freezer as soon as possible until I’m ready to process them. If you’re dealing with bulk samples then I would recommend changing the alcohol more frequently as alcohol forces the water out of the insect’s bodies, diluting the alcohol and decreasing the preservative effect. Good luck!
See, therein lies the problem, Morgan: what I WANT to do is set a bunch of traps and leave them, unattended and undisturbed for about 4 days at a time. The specimens would be collected after the 4 days and could be put into high-octane alcohol at that point… but we likely won’t have much in the way of freezer space (we’ll be in very remote areas) right away. My other issue is that 100 or 95% EtOH tend to embrittle (is that a word? If not, I’m declaring it a new word, I like it) some arthropods and cause the setae of others to fall out…all bad things for longer-term storage and especially ID. Methinks I may have to develop completely separate ADDITIONAL sampling protocol for the DNA component. Le sigh.
Bummer! I can tell you that I’ve had a hard time getting any results from traditional pan trap samples or even low EtOH% malaise traps, COI or otherwise, so if you do come up with a new technique that doesn’t allow the insects to rot in the field, I’d love to hear about it! Good luck!