CharonY

Precisely. Your job is essentially to listen and learn and ask questions if something is not clear to you. Communicate clearly and you should be fine. We generally do not have any expectations in terms experience from undergrads. And even if we had, it would be fairly unreasonable, as each lab has its own workflow. What they are going to do is to familiarize you with the inner workings of the lab and show you your responsibilities. What you should bring with you is very simple calculation abilities (for basic stuff like calculating molarities, for example) and if you are uncomfortable with it, let them double-check. In fact, at the beginning try to verify everything just to make sure that you got it (usually they will do some control, but if they are busy or overworked, they may neglect that). It is always better to ask than to ruin someone's experiments, as ecoli mentioned.

There is also the fact that studying/postdocing lasts for a few years, usually. People might get used to and subsequently get attached to their current environment. A kind of increased viscosity born from proximate familiarity, if you will. Out of convenience people may seek to avoid another change (i.e. back to their country of origin). This may even be the case if the work environment is not objectively better and I suspect that something similar is the reason why many people stay in an academic environment (despite worse job opportunities, for example).

How is a dilution from 10 ul to 5 ml a dilution of 10^-3 ? To the question, what is the total dilution of the solution that you put on the plate (note that you only use 5 ul).

Basically it is assumed that the saliva of the mosquitos triggers a weak allergic reaction. However, it should be noted that there are apparently histamine-dependend as well as histamine-independent pathways resulting in the itch. The latter mechanisms is not quite clear, but seems to play a major role.

Yes, it is just an attempt to integrate existing knowledge into a more comprehensive theoretical framework and potentially discarding or limiting prior assumptions that are not found to be universal (e.g. species concepts and similar issues).

For the actual adhesion you just need a suitable surface (such as polystyrene, though in some cases you need an existing cell layer, for example). The cells will first settle on the surface, but to actually adhere and grow you need to keep the cells happy by providing a suitable environment (including temperature, pH, and gas atmosphere), as well as nutrients. Practically you have a medium in a suitable dish and seed it with fresh cells. The cells will settle, adhere, and form a layer within that dish (if you keep them happy).

Actually fermentation does not primarily yield energy but more generally serves as a means to regenerate NAD+. Also I found it easier (and in many ways more accurate) not to see the electron accpetor of anaerobic respiration as the agent to oxidize a given substrate (an electron donor, such as glucose), as in truth many different metabolic activities yield reducing equivalents in parallel which are then used into the the same electron transport chains (with a given acceptor in the end). In terms of electron balancing it does not make much of a difference. In terms of understanding metabolite flow and the underlying reactions it does.

There are markers being associated with homosexuality. As usual with association studies not many are confirmed and linkage can be weak (e.g. Rice et al 1999, Science)

Actually, I think this is a prime example of a nomenclature being arbitrary. For instance, Glycolysis could be seen as an energy pathways under this viewpoint (it generates ATP) but also as a simple catabolic pathways funneling carbon into the TCA cycle for other, anabolic processes. The TCA cycle is not directly involved in ATP production, yet it produces reduction equivalents, which are crucial for oxidative phosphorylation . I am aware that this complicates matter, sometimes unnecessarily so. But it also highlights why biology is complicated: things are interconnected. A biochemical view tends to cut-off at a certain point in order to understand a certain sub-process (and I presume, to retain sanity). In order to understand physiological consequences of these elements their interactions and resulting emergent properties have to be understood (though we have preciously few tools and methodologies to achieve that).

Also, as an add-on note, this still could happen today (though hopefully at smaller scales), as obviously drugs can only be tested in a limited number of ways. Trials normally do not involve pregnant women, for instance (and for good reasons). And rats or other test animals do not necessarily have the same susceptibilities as humans. This is why nowadays most drugs have extra warnings in case of pregnancies. There is no 100% certainty when it comes to tinkering with biology.

The volume determines how far they can be transported through your mouth and oesophagus to your stomach, if we talk about splashes. Think about a small ingested droplet coming in contact with your mouth area, it will mostly spread out and not travel far (i.e. the viral particles will be spread nicely in your mouth area, a few may be transported to your oesophagus, but on the way there there are a lot of nice cells they can adhere to). Think of it in terms of sample loss of a sample moving through a tube or something similar with an adherent surface. Of course, if you got a huge splash and your mouth was open (thus ingesting a significant volume), then yes, that could be an additional issue (but really what have you done to do that?). But think about it the following way, if a significant amount ends up in your stomach, quite a bit will already infect your mucosal surface. I would kind of worry about that first. So even if the portion ending up in your stomach deactivates all of them, you still have a large amount of viral particles in your mucosal surfaces. Especially if the droplets are small (and hence large surface to interact). Though due to their inability to replicate (I hope) the overall health effect should still be minimal. A couple of infected cells are usually not sufficient for successful establishment of a tumor (increases lifetime risk, though). Most lentiviruses are not terribly pH stable, but it can take a little while for them to be totally deactivated (30 mins to 1h). The thing is that they are generally tested for their ability to reproduce, which in your case they should not be able to, anyway.

There are dozens of details. Mostly (as I mentioned) quantitative in nature. Certain proteins are, on average, higher expressed. Other lower. Cytoskeleton is different. They become motile again, etc. Unfortunately there is only a graduate difference, which can also be found in, say, different cell types within our body.

A decent GC/MS is quite expensive. With everything included you are looking at around 20k and up. For a standard LC-MS/MS in pharmaceutical validation we are talking more towards 50k and up (though used ones may be around for around 30k, maybe). While in my opinion I do not think that it is too hard to master (though I may be biased), I doubt that one would hire someone as a technician without at least a bachelor in a related area. Labwork and IT are for the most part extremely different skill sets. The majority of the time will be involved in creating sample prep protocols, analytical methods and validating them. The interpretation is for known compounds is fairly easy, if you are not looking into very complex matrices. There are commercial database for spectra searches (cost an arm and leg), though often just reference material is used (which is usually better, anyway, if you follow non-standard protocols). I would really figure out whether you can get training on the job, rather than trying to get one on your own. LC/MS is in principle similar, but it depends on the compounds you want to look at. I.e. can you use LC for separation or is GC more suitable (e.g. how volatile are the components to be identified). If you want to start a business (i.e. offer services for sample analysis) then you are in a world of pain of getting your system certified.

You misunderstand me. What I am saying that if you splash yourself, if it comes to viral infections it is quite likely to happen well before it actually hits the stomach acids. You have a lot of mucal surfaces that can serve as entry points. Stomach acid will only be relevant if you drink a significant amount of liquid. Droplets will just work on you epidermis and mucosal surface in mouth, nostrils. The focus on stomach acid is quite misplaced.

What you describe is basically what gazillions of groups try to do (except attaching stuff to a cell, which does not precisely make sense to me). Basic issue is that cancer cells are a part of us, hence anything that harms it, also harms us. There will only be quantitative rather than qualitative traits that we can exploit.

Well, explained. That is why I urge myriam to figure out what precisely she had as a construct. Based on the description of the events it is not clear to me whether naming it an oncogene was based actual knowledge of the nature of the construct, or not.

The potential risk lies in the precise construct. With recombination it may be possible to introduce defects or deletion into p53 which may increase the chance of the transfected cell to start multiplying. The chances are low overall, though. As a whole, other stuff that we deal with on a daily basis are much more likely to increase cancer risk than a bit of a splash.

As I said, stomach acid does little (unless you started to drink a lot of it). The worrisome interactions will well happen before that (epidermis to some point, but mostly mucous surfaces in eyes, mouth, oesophagus). As I mentioned, you should figure out what it really was. However, p53 is not an oncogene and if you had one the BSL should have been bumped to 2+-3. So I assume there is little to worry there. The only question is really what kind of mutation your construct has.

Preparative LC is not terribly tricky (well establishing good condition can be a little bit). I would disagree that the upstream steps are easy in comparison. Getting the proteins to fold correctly can be a bloody nightmare. Also, in many cases additional purification via HPLC is not necessary.

It is pretty much the same, however you have to avoid denaturing conditions. Most of the time the kits work perfectly alright (Hist-tags are pretty standard, for example). I have dozens of protocols, but they depend quite a bit on what you want to purify and the principles are much better documented in standard protocol books (which I would recommend to beginners). Check out Molecular Clonin (Sambrook) for instance.

Well, if your goal is to look at spatial distance EM, STM and AFM can be your friend.

Unless used in very specific circumstances (as a local lab lingo, if you will) they generally mean the same.

Well there is data that the physiological effects of high levels of fructose may be different than equivalent levels of sucrose (i.e. the accumulation appears to be faster as the fructose exists in the free form). But obviously the overexposure is really the big issue. Exchanging it back to sucrose would do little to improve overall health status, if consumption stays at the high levels. It has also to do with the preference of Americans for the sweet taste (and I recall a study linking it to exposure to sweetened baby food, but can't recall the reference). What is of little doubt are the health effects of high levels of fructose consumption (by whatever source). I suspect that the demonization of HFC is a bit of a marketing trick so that some (maybe even the same) manufacturer can slap "made with real cane sugar" on their product to make the consumers feel better. The only way to make it better would put less sugar into the products. But then the consumer won't like the taste...

That is quite possible and of major concern as most tox studies are not suited to identify synergistic effects (just imagine the needed throughput). That is why I am a proponent of funding mechanism-based toxicology. But that is really getting off-topic now.

C57 are pretty much a standard all-purpose strain. CBA is similar, but IIRC they (or a substrain thereof) had a lower natural incidence of leukemia and is often used in tumor research. Beside the issue with translation to human health, we are also constantly challenged by pollutants in other sources that are likely to affect our germline. Women are especially susceptible, though (for obvious reasons). So it will boil down to the question of concentration. I also want to point out that the mice were exposed to two cigarettes daily, which is quite a a bit when measured against body weight.