CharonY

With regards to autoclaving, you can do that. Just be aware that oftentimes autoclaves are not very clean and it may contaminate your solution (although it would be sterile contamination). Depends on the sensitivity of your application. If made cleanly Tris tends to be fairly stable at RT (6 months did not appear to be much of an issue), though I would check pH before use to be certain. Also note that the pH is temperature dependent. Should you put it in the fridge, let it warm up to RT (or whatever the reaction temp is) before use.

Welcome to the wonderful and expensive world of molecular biology. The moment you deal with complex mixtures, the cost skyrocket. With low complexity and well-known compounds you may get away using chromatographic methods with simple detectors.

Now that got incoherent pretty quickly. But in case you wanted to know, bacteria actually produce D-amino acids (e.g. as elements during peptidoglycan synthesis). And as I said, there are enzymes responsible for the interconversion of the amino acid from the D to the L form.

Both amino acids are found in nature, but only L-amino acids are found in proteins. Biosynthesis amino acid can lead to both forms. Transaminase are pretty much unrelated to this question, as they catalyze the exchange between amino and a keto group. Think isomerase.

As BabcockHall mentioned, the regulation is enacted via proteins, not directly via the steroids/hormones (most notably proteins of the nuclear receptor superfamiliy). There are therefore multiple layers that modulate the final expression. One is on the level of the protein with all the possibilities such as by abundance, inhibition, activation, etc. On the metabolite level inactivation of thyroid hormones (I assume we are talking T3/T4 here) there are several pathways but generally is a form of deiodination.

A bit of a tangent, but it isn't it weird to propose to ban information on things that are relatively easy to design even without a detailed manual , whereas purpose-made devices for causing harm (such as guns and ammunition) are freely available? Isn't that a bit putting the cart in front of the horse?

Whether a fragment is sticky or blunt basically depends on how the respective strands are nicked. In most cases star activity changes the sites only by a little (they still tend to be palindromic to some extent, for example) chances are that they will produce same ends as they did under normal circumstances. Due to the loss of specificity the overhang sequence may very well be different, however.

As I mentioned, I do not think that anyone has investigated the precise physics behind it in detail. The best I found was more or less a list of potential interactions with all the usual candidates (with hydrophobic interactions being near the top of the list). There are some models around, mostly from the manufacturers as e.g.: http://www.pall.com/main/oem-materials-and-devices/literature-library-details.page?id=27294.

Just wondering, how is that different from standard optical tweezers (yeah I know, I should read the articles...)?

I think the question is actually: "what is consciousness"? It may very well be an emergent property of how our brain works and does not need to be beneficial per se. Also consciousness is a physical process (as evidenced by by lesion studies). So why the distinction?

RNA is mostly present as single-strand, but it can form secondary structures with itself or other nucleic acids. Thus a certain level of denaturation is required to prevent that. The role for SDS in proteins serves several purposes, actually, of which denaturation is only one.

Great, now you made my eyes bleed.

Well, typical cyanobacteria have doubling times of a few hours...

Also smells like homework.

For simplicity in terms of cultivation I would probably go for cyanobacteria. Some have engineered growth vessels which maximize surface so that the little buggers can get sunlight very efficiently. Nonetheless, the yield for use as biofuels is still very low, even with them. It may depend very strongly on how efficiency is measure, but the conversion efficiency of sunlight to biomass via photosynthesis is closer to 2 (and lower) %, from what I have seen.

1) If you look at database entry you will generally see the complete ORF. Depending on what you need (e.g. if you also want up- and downstream sequences) you may have to switch viewers that provide genomic views and you may need to search a little bit (using the annotated genome sequence). 2) There a hundreds of websites discussing primer design. Much can depend on the type of fragment you want to amplify (e.g. length, extensions, tags etc) as well as the polymerase. But for a general primer on that topic I would just google it (e.g. http://www.premierbiosoft.com/tech_notes/PCR_Primer_Design.html) and see what applies to you.

it is usually a non-issue as the efficiency of the whole cloning experiments tends to be way lower than that of the mismatch rate. You have a slightly higher chance of creating some false products, but depending on how the designers are designed and how the cloning is going to proceed you will end up with the vast majority of the correct product for cloning. Since you have to run controls anyway, the probability of PCR purity being the culprit for failed cloning is very low. There are exceptions however, e.g. if you do some tricky amplifications with very low yields. Salts or impurities themselves are no issue at all as you do a cleanup post-PCR, anyway.

I do not think that this can be easily answered. The strength of the immune responses is determined by a multitude of factors, for example. Likewise normally the respective antibody levels will decline and can fall below detectable levels. Though it does not mean that there are no cells left that present them. In addition, antibody reactions are no necessarily 100% specific, meaning that very weak responses may be triggered with structurally similar compounds. In cases of strong reactions (e..g due to certain infections) especially IgMs can be readily detected after a year or so. More sensitive methods may even spot them longer.

Well, there are labs specialized in monitoring sleep. But with insomnia it is important to understand whether you are truly fully awake or just have a bad quality of sleep, during which you just feel awake. Sleep monitors can assist in figuring that out (or, even better, look for a specialist). There are a lot of guides and help out there to improve sleep hygiene, but it depends a lot on the reasons for the bad quality of sleep/insomnia to begin with. It could be worthwhile to checking those out, though (as following them is certainly not harmful).

One should not mix up these classes, as the impact may be quite differently. Another aspect to consider in addition to the rise of resistances (be it to either class, though far more obvious and prevalent for antibiotics), is the effect of these compounds on bacterial (and other) communities in nature. A higher prevalence of antibacterial compounds may very well disrupt certain processes in water and soil with yet unknown consequences. Finally, neither of these compounds are particularly healthy for non-bacteria, either. Granted, TCS has not shown to have any drastic harmful effects and connections with endocrine disrupting functions are spurious at best right now. Nonetheless, one has to wonder why to put it in in the first place, as it does not have any measurable health advantages.

For PCRs desalting is usually enough.

The OP was talking about a fairly standard method in molecular biology, termed blotting (specifically the fixation step after that). Blotting, in turn is a standard procedure used during Southern hybridization. I appreciate the fact that you would like to join discussions, but throwing wild assumptions around without actually understanding the topic at hand can be amusing for some readers, but especially for newcomers it may be a tad confusing. It certainly is not helpful...

Popcorn, your answer does not even begin to make sense. For the most part I would think that typical ways to immobilize DNA on nylon is blotting under alkaline conditions (the positive charges of the nylon are sufficient to retain the DNA) or cross-link with UV. Vacuum Baking is a bit more old-fashioned way and I am not even sure that the precise nature of the interaction between nylon and the nucleic acid has been well understood (or investigated). It is generally not assumed to be a covalent bond, but something involving salt bridges and hydrophobic interactions (for nitrocellulose that is; Schleicher and Schuell 1992). Which basically means that these are rough guesses.

Ammonium acetate buffers a bit higher than what you need. However, Ammonium formate should be right in your range. The slightly tricky bit is actually what molarity the prootcol demands. Many protocols actually indicates the cation portion. In that case the above approach would be appropriate (just leave ample room to fill up to final volume). Sometimes total molarity is meant, in which case it refers to ammonium formate + formate.. In that case the easiest way is to approximate with the Henderson-Hasselbalch equation (though a little bit of adjustments is often necessary, regardless).

The big issue is that there is really no path per se. There are a number of things people go through (e.g. obtaining degrees and so on) but getting a real career is incredibly hard and very volatile. The best advice is probably to go out and look what kind of jobs are actually there that would interest you and get into contact with these people. This is very tricky, but there is no educational or research path that can land you a specific position. Having a strong network of people is much more useful in the long run. Of course, you are somewhat far off and none of the crucial steps are likely to be happening right now. If you are interested in zoo work, I would try to get an (unpaid) internship there as an undergrad. That way at least you will get to know people. If you want an academic career, be advised that getting a PhD and technical knowledge is the easy part. Just as a point of reference, if you have got a PhD and some postdoc under your belt, you will be competing with about 150-250 other people with at least the same education and knowledge as you for any given faculty position . Even positions on soft money can easily get about 100 applicants. Being the perfect fit that puts you head and should above that competition is often down to luck. For instance you may have just been working on a topic that became hot just when the job was posted, but may have not been hot a few years earlier or later. Having a strong network will help to maximize your chances. That being said, you are still early in this game, and I would advise you to join a lab as early as they would take you to get a feel for the things. Realize though that what you will be doing in the lab will be the stuff you are doing up until and including part of your postdoc (i.e. post PhD but pre-faculty time). but as faculty you will see a radical shift in the things you will have to do. I also would advise you to enjoy your time, where you still got some freedom to do things of your liking.