Re: Starting with this diy stuff - Entomology
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Avery Ashley
Feb 19, 2013, 3:37:25 PM2/19/13
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Watch eBay like a hound. You'll find really great deals every now and then.
On Monday, February 18, 2013 1:13:08 AM UTC-5, Richardvs Svm wrote:
On Monday, February 18, 2013 1:13:08 AM UTC-5, Richardvs Svm wrote:
Hi! Well, I new here, and also very new to molecular biology. I took an introductory course about six months ago, and I became really interested in the field. Besides, I discovered you don't need to own the latest equipment to try some experiments.
I'm really interested in insects, and what genes they use when they face different environmental conditions, like production of heat-shock proteins or what kind of genes activate when feeding on a poisonous plant etc. There are literally millions of insects to work with... so exciting.
Oh, but I must say I own no equipment whatsoever, so, so far it's all a dream... I've read several pages, but I'm kind of lost. What should I buy first? If I wanted to check insects genes, would I need kits, or something like that?
Tristan Eversole
Feb 21, 2013, 1:02:31 AM2/21/13
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Hokay. Let me see, here….
1. You're absolutely right when you say that there's an enormous amount left to be discovered regarding the insects. The arthropods are the most diverse animal phylum (followed by the molluscs and the nematodes, IIRC), and we've described something like 1.8 million species from Class Insecta. In other words, your estimate is spot on!
2. I can give you a little advice about collecting-- this is coming from my recent experience in an entomology course. It turns out that I'm not a great entomologist, so you might want to find someone with more experience, but I hope I can at least get you started.
--The gold standard for entomology equipment seems to be that offered by BioQuip (http://www.bioquip.com/ ). They're not that cheap, but I haven't been disappointed by anything I've ordered from them. You'll need a net, a kill jar, a pinning block, a box, and some pins, at least. Insects that are too small or fragile to be mounted on pins can be kept in little vials filled with ethanol; BioQuip sells the vials too. In the class, we charged our kill jars with ethyl acetate. I think there are other things that can be used, but I'm not sure what.
--If you want to identify insects, you'll need a good set of dichotomous keys. I had xeroxes from the latest edition of DeLong's "Introduction to the Study of Insects", but it may be possible to get keys off the internet. BugGuide (http://bugguide.net/ ) and DrawWing (http://www.drawwing.org ) were both very useful (many insects can be most easily identified by wing venation), and What's That Bug? (http://www.whatsthatbug.com ) is probably pretty useful, although I've never tried asking the people there for help.
Be warned that if you intend to use the keys, you'll be looking at steps like
> 13(1'). First metasomatic segment (sometimes first 2 metasomatic segments) bearing a hump or mode and strongly differentiated from rest of metasoma (Figures 28-11, 28-83); antennae usually elbowed, at least in female, with first segment long; pronotum more or less quadrate in lateral view, usually not reaching tegulae (Figures 28-11, 28-83); often wingless.
This is the final step in the Hymenoptera key that identifies something as belonging to the Formicidae, the family that includes all ants. The terminology probably sounds horrible, but it isn't. As long as you're at the family level, identification isn't the nightmare you'd think it is. (Exceptions: moths, bees, butterflies to some degree.)
--Be aware of restrictions concerning collecting-- you may need a permit to go catching insects at a park. That said, if you can collect at a park, do. In my experience, there is an alarmingly great disparity between the insects you can find an suburban or urban environment and those you can find in a park. (There's nothing around here except cockroaches, bees, springtails, psocopterans, and, uh, earwigs. I don't understand the last one.) Surprises are quite possible, though; I found an innocellid (one of the two groups of snakeflies) randomly hanging out on a tree a few blocks from my house.
--Pan traps and Berlese funnels are both easy to build, and are almost guaranteed to provide you with something interesting.
--A stereomicroscope is really, really handy. It's probably not an actual requirement, but…. if you can get one, do.
3. I can also give you a small amount of advice about molecular biology, although other people on this list know more than I do.
--Researchers have become interested in exploring the relationship between environmental conditions and gene expression in the crustacean Daphnia pulex, in part because we know the ecology of the Daphnia genus comparatively well. D. pulex has recently been sequenced, and something like a third of its genes have no known homology. (Colbourne et al. The ecoresponsive genome of Daphnia pulex. Science (New York, NY) (2011) vol. 331 (6017) pp. 555-61. http://www.sciencemag.org/content/331/6017/555.full.html ) This is probably your best bet for studying gene-environment relationships, although D. pulex is obviously not an insect.
--However, examining gene expression in insect cells or tissues is probably not something that can be done at the amateur level.
--Basic molecular biology equipment would include a gel electrophoresis chamber, a power supply, a PCR machine, a centrifuge, a water bath, a refrigerator, and some means of imaging your gels-- I'm actually not sure what DIYers have been using to do this. (You may be able to substitute the PCR machine for the water bath.) In general, molecular biology is going to be much more expensive than collecting-- BioQuip's basic Insect Mounting Kit is $43.70 and an OpenPCR machine is $600. Kits would be the way to go; older and less proprietary protocols tend to involve things like organic solvents.
It's interesting that you mention heat-shock proteins-- they may function as "evolutionary capacitors", storing up neutral variation which is then "unleashed" by decanalization in an stressful environment. I have to admit that I'm rather skeptical, but I haven't paid much attention to the topic in a long time, and I don't know how work on the matter has turned out. I did run into a paper using morphometric methods to study wing shape variation in flies following the inhibition of Hsp90 (Debat et al. Hsp90 and the quantitative variation of wing shape in Drosophila melanogaster. Evolution (2006) vol. 60 (12) pp. 2529-38 www.flywings.org.uk/PDF%20files/Evol2006_Debat.pdf ) rather recently, though. You might like it.
--T.
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1. You're absolutely right when you say that there's an enormous amount left to be discovered regarding the insects. The arthropods are the most diverse animal phylum (followed by the molluscs and the nematodes, IIRC), and we've described something like 1.8 million species from Class Insecta. In other words, your estimate is spot on!
2. I can give you a little advice about collecting-- this is coming from my recent experience in an entomology course. It turns out that I'm not a great entomologist, so you might want to find someone with more experience, but I hope I can at least get you started.
--The gold standard for entomology equipment seems to be that offered by BioQuip (http://www.bioquip.com/ ). They're not that cheap, but I haven't been disappointed by anything I've ordered from them. You'll need a net, a kill jar, a pinning block, a box, and some pins, at least. Insects that are too small or fragile to be mounted on pins can be kept in little vials filled with ethanol; BioQuip sells the vials too. In the class, we charged our kill jars with ethyl acetate. I think there are other things that can be used, but I'm not sure what.
--If you want to identify insects, you'll need a good set of dichotomous keys. I had xeroxes from the latest edition of DeLong's "Introduction to the Study of Insects", but it may be possible to get keys off the internet. BugGuide (http://bugguide.net/ ) and DrawWing (http://www.drawwing.org ) were both very useful (many insects can be most easily identified by wing venation), and What's That Bug? (http://www.whatsthatbug.com ) is probably pretty useful, although I've never tried asking the people there for help.
Be warned that if you intend to use the keys, you'll be looking at steps like
> 13(1'). First metasomatic segment (sometimes first 2 metasomatic segments) bearing a hump or mode and strongly differentiated from rest of metasoma (Figures 28-11, 28-83); antennae usually elbowed, at least in female, with first segment long; pronotum more or less quadrate in lateral view, usually not reaching tegulae (Figures 28-11, 28-83); often wingless.
This is the final step in the Hymenoptera key that identifies something as belonging to the Formicidae, the family that includes all ants. The terminology probably sounds horrible, but it isn't. As long as you're at the family level, identification isn't the nightmare you'd think it is. (Exceptions: moths, bees, butterflies to some degree.)
--Be aware of restrictions concerning collecting-- you may need a permit to go catching insects at a park. That said, if you can collect at a park, do. In my experience, there is an alarmingly great disparity between the insects you can find an suburban or urban environment and those you can find in a park. (There's nothing around here except cockroaches, bees, springtails, psocopterans, and, uh, earwigs. I don't understand the last one.) Surprises are quite possible, though; I found an innocellid (one of the two groups of snakeflies) randomly hanging out on a tree a few blocks from my house.
--Pan traps and Berlese funnels are both easy to build, and are almost guaranteed to provide you with something interesting.
--A stereomicroscope is really, really handy. It's probably not an actual requirement, but…. if you can get one, do.
3. I can also give you a small amount of advice about molecular biology, although other people on this list know more than I do.
--Researchers have become interested in exploring the relationship between environmental conditions and gene expression in the crustacean Daphnia pulex, in part because we know the ecology of the Daphnia genus comparatively well. D. pulex has recently been sequenced, and something like a third of its genes have no known homology. (Colbourne et al. The ecoresponsive genome of Daphnia pulex. Science (New York, NY) (2011) vol. 331 (6017) pp. 555-61. http://www.sciencemag.org/content/331/6017/555.full.html ) This is probably your best bet for studying gene-environment relationships, although D. pulex is obviously not an insect.
--However, examining gene expression in insect cells or tissues is probably not something that can be done at the amateur level.
--Basic molecular biology equipment would include a gel electrophoresis chamber, a power supply, a PCR machine, a centrifuge, a water bath, a refrigerator, and some means of imaging your gels-- I'm actually not sure what DIYers have been using to do this. (You may be able to substitute the PCR machine for the water bath.) In general, molecular biology is going to be much more expensive than collecting-- BioQuip's basic Insect Mounting Kit is $43.70 and an OpenPCR machine is $600. Kits would be the way to go; older and less proprietary protocols tend to involve things like organic solvents.
It's interesting that you mention heat-shock proteins-- they may function as "evolutionary capacitors", storing up neutral variation which is then "unleashed" by decanalization in an stressful environment. I have to admit that I'm rather skeptical, but I haven't paid much attention to the topic in a long time, and I don't know how work on the matter has turned out. I did run into a paper using morphometric methods to study wing shape variation in flies following the inhibition of Hsp90 (Debat et al. Hsp90 and the quantitative variation of wing shape in Drosophila melanogaster. Evolution (2006) vol. 60 (12) pp. 2529-38 www.flywings.org.uk/PDF%20files/Evol2006_Debat.pdf ) rather recently, though. You might like it.
--T.
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