CharonY

Also, from what I understand he was instrumental in devising the strategy to begin with. Thus he is unlikely to criticize himself.

As mentioned above 10 units of restriction enzymes should be sufficient to completely digest 2 µg DNA in one hour. The only reason to add more enzyme is when the enzyme lost activity. Otherwise it won't do anything and in case of star activity it would be harmful.

The way to assess this is to know how much enzyme (in terms of activity or units, not volume) compared to the DNA. If what you describe is correct and assuming that the enzyme is not diluted and the right buffer is used you are doing a horrible overdigest anyway. Low concentrated enzymes are mostly sold with around 10 units/ml. 10 units (i.e. 1µl) is generally sufficient to digest around 2 µg of DNA within an hour under optimum conditions (depending on DNA it may even take less). You are using potentially 50 units for 2 ng. However, even if it was a typo and you used 2 µg (as 2 ng diluted in buffer would operate below the detection limit of EtBr gels) it should be more than enough. Especially as BglII had star activity, if memory serves. But more importantly, what is the rationale behind adding more enzyme? Low intensity in gels would not be a good reason.

A part of the ferritin is secreted. Also the free iron is in healthy individuals is very low but a not everything is bound to proteins. They do associate with whatever is there, so they are technically not free. But not as tightly bound as in chelators or iron scavenging proteins. The overall amount is relatively low. Blood loss anemia is the result of massive loss of red blood cells.

Indeed. One of the major jobs of science is to show what else we do not know. Sometimes we even come up with answers .

Depending on the type of uni and on the position to fill student evaluation can be an important factor in tenure reviews. Once tenured it is a bit different, of course. However, if one is not mainly a teaching prof it makes sense to simplify things regardless. The reasons is that many do not want to deal with complaining students in their office all the time. One has to keep in mind that especially in research unis teaching is of overall lower priority for the prof (though not necessarily for the uni). I have not yet heard of profs that allow cheating. What may happen though is that they do not punish "minor" cheating attempts (or overlook them). Usually that results in less hassle to deal with. I have heard that there were lawsuits regarding possible cheating attempts and especially those on tenure tracks do not want to take any risks. However many profs allow cheat sheets where one can write down equations etc. This is not too bad, as it puts less emphasis on rote learning and you test the application of the available information. But then one often cannot do too much either. If the questions do not resemble problems discussed in class the majority of the students are usually stumped, cheat sheet or not.

Get the book "Molecular cloning" from Sambrook.

You could try other filter materials for ultrafiltration (I assume you sued the amicon columns and you used the correct MWCO?). Otherwise you could make a cleanup using affinity columns (often sold as desalting columns). However I am not sure whether they are really much cheaper than a small dialysis tube. In fact, I think they are roughly in the same price range. Why is the quantity of solvents limiting? Or you could precipitate the protein and wash the pellet to remove buffer and salts (though some proteins do not easily resolubilize).

It is basically the same in university. The thing is that not only the system promotes it, but the students, too. Teachers that try to focus on a system in which the understanding is tested generally get low student evaluations as they consider it too hard, non-intuitive etc. Of course it is not necessarily the case in all classes but on average as a teacher you are better off if you e.g. show when to use which equation and then simply test for their correct application rather than asking complex questions.

Depends on which regions change. If the binding to the surface receptor of the host gets diminished in the process, then yes.

Most polymerases have a proof reading element, which the HIV polymerase lacks. As mentioned, it already lacks other DNA repair mechanisms Note that errors are everywhere not only in specific proteins. However, those that are defective are simply not infectious and those that do not provide any selective advantages (as the receptors) vanish or are not detected. As viruses do not have to maintain metabolism and reproduce in huge quantities, it can allow itself to produce a lot of crap.

By definition anything that is supernatural is outside the realm of science. If it was testable, it would suddenly not be supernatural anymore.

Interestingly enough, for many if not most toxic compounds only vague mechanisms are known. There are few for which all the involved pathways are elucidated. The reason is that many have unspecific effects. There is hope that systems biology approaches may be useful in identifying novel toxicity pathways, though.

As a rule of thumb there is quite some research needed to establish potential adverse effects of any given GMO strain. As extreme effects are usually rather unlikely the difficulty is of course to identify effects that are above background toxicity (and how to define that baseline). For instance GMO maize may have more negative effects than pristine WT maize, but have less or equivalent to those grown under normal agriculturally relevant conditions (including use of pesticides, environmental pollution etc.) Unfortunately biochemistry will not be the ultimate answer either. Even if toxicity pathways are identified we will be stuck with BMD or similar parameters that do not necessarily apply to humans. But that is a current limitation of traditional toxicology.

That is incredibly vague and depends a lot of the nature and use of the construct. To answer in an equally vague fashion: First step is often PCR (or several PCR steps for specific constructs) followed by cloning.

Error prone replications system.

While it is not precisely news, the paper makes provides compelling arguments that are especially of interest for biologists. Essentially because it makes an intriguing argument by discussing it in the context of an important part of human societies. It is simply one of the more elegant and easy to read articles about this topic that I came across. In fact, one could post it as easily in biology, philosophy as well as ethics as it touches all these elements. More to the point, the author also offers compelling arguments against stochastic effects as part of free will. The lack of predictive abilities does not per se lead to the conclusion that there may be a free will. Actions may not be determinable or even deterministic, however, it neither means that the individual in question had a the possibility to behave differently. The discussion in the light of the justice system is of special interest. Also, from the biological side it is obvious that so far there is no mechanism or even theory that could account for something like free will. However much of our society is built around that concept.

I found a very interesting assay regarding free will in PNAS. The abstract: And a paragraph from the conclusions: While not radically new, the paper is very good read and also includes implications for the justice system (also note the follow-ups). It is an open access article: http://www.pnas.org/content/107/10/4499.abstract

One would have to determine precisely whether it is the same compound (unfortunately I forgot the name), however for the one in question was untested, and hence the reluctance to use it. IIRC it was not submitted for toxicity analysis and I would be surprised if it was used in anything more than small scale. And of course the benefit of it in relation to risk has not been proven for that scale either. The analysis of toxicity is not trivial, especially the evaluation of ecotoxicity. Edit the nanotech I was thinking about is from a company called green earth technologies. Based on what I know about nanotechnology their claims are highly dubious, especially the proposed micelle size (but then it is hard to evaluate as they do not say what it is). Is it the same you are thinking of? That product was only released in April, and has therefore never been tested in the field afaik.

I think a rabbi and a catholic priest are missing from the picture.

You got your definitions all mixed up. The epitope is a part of the antigen. Also antibodies can be antigens for other antibodies.

Well actually I would turn the table and ask you what kind of career you aspire to. Then we could tell you if a given career track (as far as we know) may align with that. Real jobs and often even academic jobs are not necessarily locked down on a topic.

There are a number of mitochondria-related diseases. However in case of lethal mutations (or anything that abolishes reproductive abilities) they obviously cannot spread through the population.

Predicted does not mean that the sequence is predicted (it has been sequenced) but that it has been predicted to be an open reading frame. This topic should be covered by most molecular genetics text books (e.g. Genes). Again, the function of a locus is predicted but the sequence itself is based on data (though depending on source it may be faulty, but that is another issue).

I think I know the proposed stuff. If it is the same as I was thinking of then it is some kind of nanomaterial with unknown formulation. The EPA requested the composition which the manufacturer denied to disclose. So far there is only the word of the manufacturer that it is harmless. Considering the fact that nanotoxicology is still quite in its infancy I am not sure whether it would be a good idea to throw that stuff in. For instance it could increase the solubility and hence, bioavailability of the oil. Well and most of the offers from allies during the aftermath of Katrina were declined, too. http://www.washingtonpost.com/wp-dyn/content/article/2007/04/28/AR2007042801113.html Despite waiving the Jones act, unfortunately.