hi adam, just testing. did you get an email?
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Equipment for field sampling
- autoclaved screw-top vials with moist paper (if sampling live beetles)
- autoclaved empty screw-top vials
- squirt bottle (or separate vials with ethanol)
- bullet box for vials
- larger tubes
- chainsaw + gas + oil
- 2 broad chisels
- crow bar
- anvil pruner
- folding saw
- soft forceps on a string
- hard forceps
- scalpel blades
- lighter (for forceps & scalpel sterilization)
- ziploc bags
- permanent markers (alcohol-resistant)
- pigma pen for labels
In the field
- beetles: pierce head, put in numbered vial, record
- larvae & frass: put in numbered vial or tube, record
- fungi: same as above
- frontalis: small samples tied up with rubber band, in ziploc bag
Sampling bacteria from beetles
Up to 3 larval chambers, and up to 3 adult beetles (ideally new generation) per tree
1. surface wash 2. the beetle crushed (elytra removed, surface-washed in bleach, water, ethanol, and dried out)
Materials for dissection
- eppendorf vials
- 1000 uL pipette tips
- EtOH bath
- vial rack
- surface wash for adult beetles (elytra removed, surface-washed in bleach, water, ethanol, and dried out)
Experiment with SDS-EDTA.
- Please make 100 ml solution of 2% SDS and 100mM EDTA. Borrow these chemicals from the Smith lab (Adam Black know that you are coming) and use deionized water.
- Prepare 18 live beetles from one of the plastic boxes in the lab.
- Pour the solution in a small soda bottle, and throw 9 beetles in it. Do not close the bottle! Hang the bottle (open) outside on a tree where no one will disturb it.
- The other 9 beetles will go in ethanol and in the freezer; label the tube “SDS-EDTA experiment”.
- 5, 10 and 15 days from the start, remove 3 beetles from each treatment, extract DNA (MoBio kit) and run a gel. Measure DNA concentration using quantitative ladder.
Put together a selection of common bark and ambrosia species from this area (pinned). No more to 3 specimens per species, no more than 10 or 15 species total. This is for our international collaborators – they will send us specimens from their regions.