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Author Topic: Need germ-geek help
ScholasticSpastic
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Icon 1 posted January 26, 2007 10:18      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
Here's a rundown of what I'm doing for back-ground:

Soil cores were collected from various parts of a wetland with high Se levels. We've also taken water samples to test for Se- our eventual goal is to see if there's some correlation btwn soil fauna and Se levels. The cores were homogenized (bag squishing) and sub-samples of 10g were isolated from which all cultures will be taken.

1g portions of the subsamples were washed with distilled water and serial dilutions followed up to 6 log ten. We used a media broth for the serial dilutions to avoid causing undue osmotic stress to the wee bugs. Each dilution from each site was plated out on King B media (total 49 plates + two controls and a christmas-tree from my tongue fauna for s+g).

From these plates I then picked 6-11 colonies that I felt represented the variety found thereon and streaked these out on their own plates. Prior to this we had Christmas break (hence the tongue christmas tree) and all plates were refrigerated to slow growth so I still had some edges to streak from.

Here's the problem: The plates I've streaked out for homogenous colonization have a HUGE rate of failure (ie no colonies). Only 12/58 of my latest plates show signs of colonization from the parent colonies after four days.

Culture conditions: King B media (makes soil pseudomonads look yellower) @ 25C (we just leave them out on a counter-top) for 4-7 days.

My personal hypothesis is that the refrigeration and long wait between growing out the parent colonies and streaking out singles has negatively impacted the viability of the colonies I've picked.

What I'm looking for here is a load of other hypotheses regarding my failure so I can test those, too. We need to get these bugs grown out and sequenced so we can identify them.

(We're doing 16srDNA PCR magnification and then comparing sequence to a database to identify at least genus if not species. Next, we're going to repeat the process using a soil kit that automatically collects DNA from a soil sample, then we're going to make E. coli plasmid libraries to eliminate noise and have THAT sequenced so we can compare the results of the different protocols. Then- joy!- we may start looking at the rhizospheric fauna!)

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

Posts: 540 | From: Vernal, UT | Registered: Jan 2007  |  IP: Logged
nerdwithnofriends
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Icon 1 posted January 26, 2007 11:36      Profile for nerdwithnofriends     Send New Private Message       Edit/Delete Post   Reply With Quote 
You know, when I try to talk to other intelligent people about my occupation and interests, often times they'll kind of understand, but not quite. I've never been able to sympathize with them, until now.

I'm not a microbiologist, but my old man is (was), so all I can suggest is maybe you need to change the environmental factors. The one that comes to mind most readily is the oxygen available: are these anaerobes, or not? Maybe cutting off the oxygen would help them to grow.

I'm pretty much talking out of my ass here, so I'll shut up and let the members who know something talk. Doesn't MMKK do some kind of environmental microbiology or something?

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"The Buddha, the Godhead, resides quite as comfortably in the circuits of a digital computer or the gears of a cycle transmission as he does at the top of a mountain or in the petals of a flower." - Robert M. Pirsig

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Xanthine

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Icon 1 posted January 26, 2007 12:25      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
Well, the laws of the universe don't change, so if they grew on your benchtop before they should do it again. Unless, of course, there's something wrong with your cells.

business attire is into medical microbiology. Where is she? She might know a trick or two.

Unfortunatley, I'm more interested in the bits and pieces of germs than the germs themselves, so I mostly just clone gene fragments into E. coli. Which means I'm pretty good at growing E. coli (which is like saying you can successfully breed rabbits [Razz] ), but not much else. That said, the lore in our lab is, if you leave your plates in the fridge for more than a couple days, they are basically toast.

It's possible they sporulated, but my understanding of sporulation is, once the bugs have returned to a happy environment, they start growing again.

Another thought: is your media still good? How are you spreading the plates? If you're using a loop, are you sure it's completely cooled off? If you're using a glass spreader that you sterilize with burning alcohol, are you sure the ethanol is all burned off and the spreader is cooled? If you're a toothpicks kind of guy, are your toothpick nice and clean? If you use beads, are they clean and dry? These are stupid ass questions, but that's the best I can come up with. I had a nightmare couple of weeks last spring when I couldn't get my cultures to grow in liquid media and I discovered that the problem was in our reusable (and very much re-used) glass culture tubes. We'd wash 'em out with detergent, rinse them, and sterilize them, but we'd gotten sloppy about the rinse and there was detergent residue in the tubes that was poisoning the cultures.

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

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littlefish
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Icon 1 posted January 26, 2007 12:36      Profile for littlefish   Author's Homepage     Send New Private Message       Edit/Delete Post   Reply With Quote 
I have a friend who worked with bugs. Unfortunately they got a contamination and had to sterilize the entire lab - washed everything with alcohol.

What I learnt from him was that organisms are temperamental. And you can make pretty much anything glow if you want. (he worked with the FPs - GFP, YFP, BFP)

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Xanthine

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Icon 1 posted January 26, 2007 23:58      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
We have contamination issues too, especially in the spring and fall. For some reason, salmonella really likes to bloom in spring and fall, and it's got some resistance to the antibiotic we use as a selectivity marker (doubling the drug dose can help, but the cells grow slower). One year it got so bad I wanted to just burn my entire bench, but my boss wouldn't let me, so I threw out all my buffers and doused everything, including my burner, in 10% bleach and followed up with alcohol. That helped for a couple weeks and then I had to do it again. We spike most of our reagents with sodium azide, sterilize them either by autoclaving or filtration, and refrigerate the ones that really tend to grow things, but you can't put azide in a cell culture. And, sometimes, even the reagents spiked with azide grow things.

I'd really love to know what it is that grows in our 1 M MgCl2 stock. Seriously. We make our solutions with ultrapure water, so I know there is nothing in there except water and MgCl2. No carbon. No nitrogen. What the hell is that bug eating??

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

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ScholasticSpastic
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Icon 1 posted January 27, 2007 00:30      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
quote:
Xanthine said:
Well, the laws of the universe don't change, so if they grew on your benchtop before they should do it again.

I agree. The reason we're culturing these bugs on the benchtop is that some of them would be happy in the fridge and others would be happy in the incubator, but everyone is likely to be happy at room temperature (or at least not too unhappy). Also, I've picked the colonies that originally responded at room temperature. I would be surprised (but don't discount) if the period of refrigeration toasted them- this is a very temperate climate (our high today was 12F). I also agree with Xanthine's thought that sporulated bugs would begin growing again when culture conditions were optimal (or not deadly) again. Then again, I'm not a microbiologist either, I'm just a sophomore playing with sophisticated toys. I look forward to taking a micro class, though!
quote:
Nerdwithnofriends said:
You know, when I try to talk to other intelligent people about my occupation and interests, often times they'll kind of understand, but not quite. I've never been able to sympathize with them, until now.

Perhaps you could feel that way if this were my occupation, but I'm lost, too. I speak with authority about everything and I speak from experience about nothing [Geek] .

quote:
Xanthine said:
Unfortunatley, I'm more interested in the bits and pieces of germs than the germs themselves, so I mostly just clone gene fragments into E. coli. Which means I'm pretty good at growing E. coli (which is like saying you can successfully breed rabbits ), but not much else.

We'll be doing that next, actually, as part of our fun with the soil DNA kit. If we don't insert our 16srDNA fragments into plasmids they'll just be a bunch of noise when we have then sequenced. We have the tools to sequence DNA in our lab, but the process is sufficiently tedious without automated equipments and <joy!> robots that I've allowed my professor to talk me out of it. We'll pay to have someone else get the code and then we'll do the database comparrison ourselves.

Also, Xanthine, (and these were all good ideas) the loop was cooled (I doused it on a sterile plate to be sure I wasn't cooking the bugs when I picked them- there were no cfus on my control plate, so the odds are good that I didn't contaminate too many plates by doing that), the toothpicks were autoclaved (now I've decided that I prefer the toothpicks), the glass spreader was only used with aqueous/broth aliquots and they all look great [Applause] , and the petri dishes I use are, of course, single-use plastic trash (but they're sterile trash, so that's okay). What are these beads of which you speak? I'm intrigued... Oh, also, the media was fresh (I make all my own media from a very simple recipe- I'd question the media, but it's the same thing I used to grow the parent colonies).

quote:
Nerdwithnofriends said:
The one that comes to mind most readily is the oxygen available: are these anaerobes, or not? Maybe cutting off the oxygen would help them to grow.

Thanks, and I thought about anaerobes (because I felt that anaerobic bacteria were more likely to play a hand in metabolizing bioavailable Se compounds as Se is very much like sulfur), but my professor convinced me that we should stick to critters that will culture without such specialized requirements. Most soil bacteria are facultative anaerobes, which means that they do well in the absence of oxygen, but they're also fine in its presence. E. coli is that way, too, Xanthine! Also, it'll metabolize bioavailable selenium. (I'm learning to appreciate E. coli for the superior species that it is.)

I want to thank you for responding! Mostly what I'm interested in is the brainstorming of those who aren't involved in the project and thus don't have our protocol blinders obscuring their view. There are no remedial or useless suggestions because I'm not quizzing you- I'm stumped! If I haven't tried your suggestion yet (or if you don't know I've already tried it) then I'd be a real mule to accuse any of you of being unhelpful.

P.S. Please forgive any spelling errors that arrise from inclusions or exclusions of doubled letters, I've always had a tough time with that part of our language.

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

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ScholasticSpastic
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Icon 1 posted January 27, 2007 00:35      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
quote:
Xanthine said:
Seriously. We make our solutions with ultrapure water, so I know there is nothing in there except water and MgCl2. No carbon. No nitrogen. What the hell is that bug eating??

I saw a really cool documentary once in which a film of subterranean bacteria was able to obtain biochemical energy from a solute gradient in an underground cave. No nutrients, just the little bit that they were able to scrape together from the remains of their ancestors. No energy for them but what they could salvage from the different ion densities in their water column. Bacteria will grow. Period. (They Rock!)

Edit: is this distilled and deionized H20 in which they're growing? You can miss a lot with just distillation. Have you tried autoclaving the liquid fraction of your solution before you drop in the MgCl2? Is this an old dry stock of MgCl2 that might be harboring unseen visitors? Or do you also get opportunistic colonizers from factory-fresh MgCl2? Have you tried autoclaving the 1 M MgCl2 solution after mixing it? It's such a simple solution that I can't think of how it would be harmed by high temperatures.

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

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Xanthine

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Icon 1 posted January 27, 2007 01:02      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
We use deionized water that's been passed through a Milli-Q. We're a crystallography lab. We like to know exactly what's in every solution. No mystery ions allowed. But that is a good thought about something coming along for a ride in the solid Mg. It's a salt we use a lot of so our stocks aren't that old, but the air is full of crap here. Thing is, while we tend to be a little too impatient and lazy to autoclave our solutions, we do filter-sterilize them. That should keep the bugs out, and it certainly takes the dust out (dust is not my friend). And shit still grows...maybe it came in on a bottle or someone's pipet tip (but we autoclave those...then again our aseptic technique ranges from virtually nonexistent to freakishly good).

I did know wild E. coli can grow anaerobically. Our lab strains aren't very good at it, but lab E. coli is basically a domesticated bacteria and it needs some coddling. I didn't know E. coli can eat Se, but it makes sense. People can and do feed E. coli selemethionine to make proteins labeled with Se. Growing the bugs in the presence of selenomet works quite well. Getting the labeled protein to cooperate is another story sometimes, but that's not the bacteria's fault!

Come on, you want to do the Sanger method. It'll put hair on your chest and scars on your soul...and when else will you get a chance to coat your shoes and your pants in acrylamide?

One last edit for I turn in: the beads I speak of are large glass beads used to spread a lawn on plates. Not very useful for you right now I take it, but basically what you do is squirt some liquid culture on a plate, add a few of these sterilized beads, and gently shake your plate on the benchtop for a few seconds. Dump the beads in a beaker for future washing, drying, and autoclaving and put the plate away to incubate. Very easy. Much better than the old-fashioned spreaders with flaming ethanol. We keep our sterile beads in ordinary glass bottles, and, like I said, they're completely reusable. Our main source of turn-over is morons like me dropping them all over the floor and losing them.

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

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ScholasticSpastic
Highlie
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Icon 1 posted January 27, 2007 07:52      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
Xanthine,

Look at you go, you biochemist, you! Kicking out that terminology that underscores just how much I still have to learn... Making me look up Sanger method to see that that's the name for what I was talking about...

Selenocysteine is another good amino acid in which a selenium atom is substituted for sulfur. I haven't memorized all my amino acids yet, but that's one of my goals for this semester.

Difficulties with Selenomethionine cooperation probably arrise from the larger atomic radius of the Se. It can adversely effect tertiary and quaternary folding of proteins and if you've replaced a sulfur that participates in a sulfur bond you'll get a weaker bond from the Se. I suspect that's part of how selenium toxicity is bad (m'kay?)- we get kinky proteins. But, again, I'm just a sophomore and I'm very likely wrong.

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

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Xanthine

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Icon 1 posted January 27, 2007 09:22      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
Sorry man, I didn't mean to offend, it's just that after keeping a lid on it for years I just got excited!

But now you learned why sane people send out for sequencing, and rich people buy the machines. [Razz] Actually, it's all the Sanger method, but there's the cheap, old-fashioned, radioactive and tedious Sanger method and then there's the fast and easy, let's-have-a-computer-do-it-for-me Sanger method. The only people who choose to do the latter are...well, I actaully don't know anyone who, given the option, would choose to do it the old-fashioned way. Except for ths one whack prof at my old university, but did I mention he was whack?

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

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ScholasticSpastic
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Icon 1 posted January 27, 2007 14:26      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
Xanthine, it takes more than an exposure of my ignorance to offend me: I really didn't know it was called the Sanger method. I just knew it was mo' betta' to let someone else take care of that part for me. You will learn that I am offensive far more often than I'm offended [Razz] . Any discussion of this nature is welcome and appreciated- the natives out here are ignorant savages and there's only so much time I can spend monopolizing my professors' time.

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

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Xanthine

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Icon 1 posted January 27, 2007 15:05      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
You could do what I used to do and harass an unsuspecting grad student or post-doc (we only pretend to be ignorant savages). Of course, now that I'm a grad student, I sometimes want to slap my undergrad, but I guess that's my punishment for the grey hairs I gave the post-doc I used to work with.

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

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ScholasticSpastic
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Icon 1 posted January 27, 2007 16:00      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
I would have been a graduate student if I hadn't allowed myself to be distracted by stupidity. We don't have graduate students out here- only four-year programs are available. My plan is to get as close to a 4.0 as I can and flee elsewhere after my Senior year. I just blew my 4.0 last semester, but they tell me a 3.91 is good, too. Myself, I'm not so sure- I'm thinking about retaking Calculus. I earned my B, but I don't have to be happy about it... Our biology department here at the Utah State University Regional Campus (Vernal) consists of four professors and maybe a total of twenty Bio majors. Most of our Bio majors are going on to Dental school or seeking work with their BS through the Forrest Service. I may be the only one right now who intends to pursue a career in science. Our Biology department is supported by a healthy load of nursing students, but you'd be surprised at how many future medical professionals seem to be utterly disinterested in bacteria.

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

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Stibbons
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Icon 1 posted January 28, 2007 00:34      Profile for Stibbons   Author's Homepage     Send New Private Message       Edit/Delete Post   Reply With Quote 
quote:
Originally posted by ScholasticSpastic:
Our Biology department is supported by a healthy load of nursing students, but you'd be surprised at how many future medical professionals seem to be utterly disinterested in bacteria.

[Insert "nurses != medical professionals" joke here] [Razz]
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Xanthine

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Icon 1 posted January 28, 2007 09:01      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
Stibbons, in the US healthcare system, you usually have to wade through several ranks of nurses before you're allowed to see a doctor, but that's okay, because the nurses actually take care of you. The doctors just look at charts.

I find that distressing, SS, just like I found the number of pre-meds at my old university who claimed to hate all forms of chemistry distressing.

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

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ScholasticSpastic
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Icon 1 posted January 28, 2007 11:32      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
quote:
Xanthine said:
I find that distressing, SS, just like I found the number of pre-meds at my old university who claimed to hate all forms of chemistry distressing.

Whoa, man (er, woman), do you mean to say that drugs are, like, some sort of chemistry? That's, like, totally messed up!

I'm frustrated by the number of people in a great many scientific and health-related fields who feel they can do their job without any real understanding of chemistry. What's the deal with Biologists who think they can get away with not understanding chemistry? Is all life not a collection of interdependent chemical reactions? I'm astounded that there are even Biochemists who seem not to get some of the basic principles that I'm learning in O-Chem right now. I understand that we live in an era in which it is impossible to be a true renaissance (wo)man, but chemistry is central to so many fields that there aren't very many scientists who, I feel, should be allowed to get away with ignoring it. (Maybe- and this is a big maybe- Astronomers don't need to know organic chemistry, but they'd better know all about hydrogen!) If a BS in chemistry were allowed at my regional campus I would totally (!!!) be working on a double-major right now.

On an unrelated subject (and I am not offended), could we aspire to abbreviating my nickname without the SS? I am not a holocaust denier and so I can't help but feel some (shiver) trepidation about the SS appellation. We've got a Spaz in the forum, but I think I may still be the only Spastic. So maybe you could just leave the Scholastic part off. Or, if you like, you can call me something else that you make up and just let me know so I can respond when you call me by it.

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

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Xanthine

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Icon 1 posted January 28, 2007 18:18      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
quote:
Originally posted by ScholasticSpastic:
]Whoa, man (er, woman),

That's okay, even people who really ought to know better get that one confused from time to time.

quote:
I'm astounded that there are even Biochemists who seem not to get some of the basic principles that I'm learning in O-Chem right now.
Oh believe me, it's worse than you know. I once had a mighty row with a labmate who's two years ahead of me concerning basic thermodynamics that everyone covers in freshman gen chem! There's two things going on. First of all, a lot of people learn just enough orgo to pass the class and then purge their brains. Even those of us that didn't are rusty through lack of practice (give me enough time and I can draw some intelligble mechanisms, but, since enzymology isn't related to what I do, reaction mechanism aren't something I think about much). The other issue is a lot universities tend to approach biochem as chemistry lite, and so all those students that can't hack the full calorie chemistry courses partition into biochem. Of course, there are exceptions. I couldn't decide what I liked better, biology or chemistry. I ended up majoring in biochem and minoring in chem (only way I'd ever finish in the four years my scholarships allowed), but I could have just as easily flipped that around. Except I didn't want to take the advanced chemistry labs because I was/am such a klutz I was afraid I'd injure myself in the name of science, and that's not as admirable as it sounds. Especially in an undergrad lab course, where the results aren't exactly news.

The trouble biochemists who thought they were getting chemistry lite face is, when and if they go on to grad school, they will find themselves being taught and advised and examined by professors who are very solidly grounded in full calorie chemistry, and they get their asses kicked so hard that years later they are still bruised. I saw it happen to some of my peers.

Mr. Spastic okay?

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

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ScholasticSpastic
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Icon 1 posted January 28, 2007 21:41      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
quote:
Xanthine:
Mr. Spastic okay?

Well, Mr. is an honorific... I'm not sure I rate one of those. But I'm fine with it as long as you understand that the Mr. part is entirely optional. Definitely not ever a Miss. When/if I get my Ph.D. I'm going to make everyone call me Dr., but until then I am only an egg.

The whole reason I'm working in the Chemistry Help Center at my campus is that I'm trying really hard to avoid a memory purge of my own. It hurt too much gaining that information to let it fade without a fight. Also, working on-campus allows me to avoid sinking back into the retail hell (I HATE it when my friendliness must be feigned).

I'm also a klutz. It's almost comical watching me try to run an electrophoresis gel. Almost... I'm doing wrist and hand strengthening exercises to try to take care of the shakiness.

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

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Ugh, MightyClub
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Icon 1 posted January 29, 2007 10:09      Profile for Ugh, MightyClub     Send New Private Message       Edit/Delete Post   Reply With Quote 
quote:
Originally posted by Xanthine:
Except for ths one whack prof at my old university, but did I mention he was whack?

Xanthine, you must be confused. There were no whack professors at UR. [Big Grin]

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Ugh!

Posts: 1742 | From: Ithaca, NY | Registered: Dec 2004  |  IP: Logged
Xanthine

Solid Nitrozanium SuperFan!
Member # 736

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Icon 1 posted January 29, 2007 10:25      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
Didn't spend much time over in Hutch or at the Med Center, did you?

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

Posts: 7670 | From: the lab | Registered: Mar 2001  |  IP: Logged
Ugh, MightyClub
BlabberMouth, the Next Generation
Member # 3112

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Icon 1 posted January 29, 2007 11:48      Profile for Ugh, MightyClub     Send New Private Message       Edit/Delete Post   Reply With Quote 
I'm of the opinion there is a "whack inversion field" surrounding that school. The normal profs are the ones that stand out [Wink]

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Ugh!

Posts: 1742 | From: Ithaca, NY | Registered: Dec 2004  |  IP: Logged
Xanthine

Solid Nitrozanium SuperFan!
Member # 736

Member Rated:
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Icon 1 posted January 29, 2007 11:51      Profile for Xanthine     Send New Private Message       Edit/Delete Post   Reply With Quote 
/me thinks back

I concede the point. [Razz]

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And it's one, two, three / On the wrong side of the lee / What were you meant for? / What were you meant for?
- The Decemberists

Posts: 7670 | From: the lab | Registered: Mar 2001  |  IP: Logged
Colonel Panic
BlabberMouth, the Next Generation
Member # 1200

Icon 1 posted January 29, 2007 17:47      Profile for Colonel Panic         Edit/Delete Post   Reply With Quote 
Maybe your colonies are nudist colonies.

Nudist colonies don't take to refrigeration well at all!

Colonel Panic

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Free! Free at last!

Posts: 1809 | From: Glacier Melt, USA | Registered: Mar 2002  |  IP: Logged
Colonel Panic
BlabberMouth, the Next Generation
Member # 1200

Icon 1 posted January 29, 2007 18:19      Profile for Colonel Panic         Edit/Delete Post   Reply With Quote 
Spastic,

The real genius in science is the ability to take something very complex and make it seem simple. It's not unlike solving a quadratic equation.

Allow me to point out long-term for a moment. I know from experience that the N.I.H. is not going to hand you over a grant simply because you have the ability to make an experiment about swamp water seem like brain surgery. They don't give rat's ass about how smart you sound; they want answers to problems -- ones a scientist first has to help them understand exist.

Now a little bit more short term, I you ever make it to the point where you have to defend a Doctoral Thesis your commitee will bust your gonads until you understand the true meaning of the word humility. First you have to make a point, then you need to let them understand that you are but a humble speck in a giant universe of the unknown.

As for your experiment, it seems like sloppy technique or a case of too many variables has helped kill your "beasties."

Don't be too disheartened, if you continue to kill your cultures, you may have a bright future as an autoclave.

Colonel Panic

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Free! Free at last!

Posts: 1809 | From: Glacier Melt, USA | Registered: Mar 2002  |  IP: Logged
ScholasticSpastic
Highlie
Member # 6919

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Icon 1 posted January 29, 2007 19:22      Profile for ScholasticSpastic     Send New Private Message       Edit/Delete Post   Reply With Quote 
Colonel Panic, You're right. I'd make one hell of an autoclave: lots of pressure and full of hot air. I also agree that there are probably too many variables and that the technique is a bit sloppy. Otherwise, I'd have to be upset that you would imply such a thing!

The trouble is that I'm new at this whole culturing thing and that my professor is a biochemist (not really a microbiologist) and so she's not the font of information she might otherwise be. I'd be having a much better time if I had taken micro, but I've been too busy taking other classes and I won't get around to it until next year. In my defense: the professor I'm working with has actually shot down a few of my ideas to control variables.

I've got another set of plates growing from the same sub-samples, but I was hoping to have a short-cut handed to me by someone more experienced than I am. These new plates will not be refrigerated and we'll see if I can get some better growth from them.

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"As in repeating a well-known song, so in instincts, one action follows another by a sort of rhythm; if a person be interrupted in a song, or in repeating anything by rote, he is generally forced to go back to recover the habitual train of thought..." (Darwin, The Origin of Species)

Posts: 540 | From: Vernal, UT | Registered: Jan 2007  |  IP: Logged


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