CharonY

Fertilizers are probably less worrisome in terms of toxicity. However think about the ecological impact. E.g. algae growth, sudden biomass increase etc.

For the most part correct. The phosphorylation is indeed done to ensure that the vector really carries an insert. Of course, doing it alone does not yield much info, but it does if compared to other results. Most controls only do make sense in comparison with another experiment. So if you compare b to c what outcome do you expect (in terms of colonies), assuming all steps of the cloning experiment worked perfectly. Under which condition would that not be the case? Visualize all the steps of the experiments and then speculate.

Nope. The microRNA is part of the RISC complex. Together they they act on mRNA.

For transformation frequency the same applies. The uncut plasmid will always have a higher frequency than any ligation. How would that be a control. What info would that give you regarding your ligation?) Again, generally it is a control for the cells. In addition you can check ligation efficiency. Regarding the dephosphorylation: you are right that it prevents self-ligation. But think about it a little more. What effect would that have on the cloning? I.e. why do you dephosphorylate in the first place? Following that line of though, what would you expect when comparing b and c and what would it tell you?

If you really want to do lab work in the bioengneering area you could try to either get a technician job after either bachelor or, probably more preferable, a masters. The big question, however, is where you want to work. While genetic manipulation is all nice and good most of the time you will also need e.g. solid fermentation or similar techniques. In the end it is important to inform yourself about what work you want to do and plan your curricula appropriately.

If the pI is lower than the ph of the buffer they will be negatively charged, yes.

a) is not for finding the efficiency of the transformation, as it will have to evaluated for each construct differently. This is because the transformation efficiency is dependent on the cells as well as the construct. However it is a control for the competence of your cells. For b and c, what do you think will be the difference between those two conditions. What does the phosphatase do and how does it affect the ligation?

Moved to the homework area. Also it would help if you would just shortly copy the question in here instead of using a link. Spammers are everywhere.

Antioxidants are old news, too.

You should check whether the effect is reproducible. During evaporation CO2 and hence, buffer capacity gets lost. However, the proteins themselves are afaik not different enough to result in different pH (I would have to check the sequence to verify that, though). I would rather assume that it is essentially a problem of a non-buffered solution. Small difference of additives or even protein concentration can shift the pH easily.

There are a number of mechanisms and the majority is probably still unknown in detail. Most rely on certain gradient e.g. along developmental axes. One well-studied element is the chromatin state of Hox genes. See for instance: Howard Y. Chang, et al. Science 326, 1206 (2009), for a relatively easy to understand read on the topic.

Sustainable is missing somewhere!

Quantum solar organo femto-crystal dots.

Well evolutionary models can be derived on the gene level, which is done routinely. However, if you want to make population studies and derive for instance the chance of fixation, you will have to figure in fitness. The latter can only be achieved by measuring on the level of reproductive units, which generally means the whole organism. Even mobile genetic elements, which are as individual as possible when it comes to genes need a vehicle to persist through generations. Though of course the horizontal transfer can be measured with other metrics.

The single most important bit of advice is not to focus too much on the courses per se. In reality you will most likely already be a grad student before realizing what benefits you most. In many if not all cases there is no clear trajectory in which you can project where you will end up and what precisely you will need for a given project. A good foundation in all the basics is helpful, of course, but chances are, you will have forgotten a lot once you come to the point where you actually seriously need it. Having a good basis makes it easier to catch up, though. Finally the most important bit is to realize that grad school is not an end to itself. You will ask yourself where do you want to be in the end? Academia? Why? Or industry? Why? What other alternative? The PhD can be an entry ticket for something, but unless you just study pro arte, you should be sure that you are moving in the right direction.

Good to hear. And honestly, it is still early days. Generally you are doomed once you get your PhD (unless you made realistic career plans way ahead of that, which most don't).

If you want to discuss this, maybe you should start a new thread. There are a couple of things in this thread that are somewhat inaccurate, but I do not want to revive arguments from two years back.

Yes, indeed, they can be intermediates of amino acid degradation. Many bacteria have a more efficient way of utilizing amino acids, though. Instead of the Ehrlich reactions (and ending with alcohols) they go back via coA dervatives to fuel the TCA. Essentially in those that utilize amino acids also as C-source.

So let's see. Your problem is the connections of glucose and pyruvate? Well how do you go from glucose to pyruvate (essentially you answered it in your post yourself). Now pyruvate is well connected to other pathways. What is the central hub in biochemical pathways in almost all organisms? From there it is a relatively small step to find the intermediates to go all the way down to the various alcohols.

Probably you should read up on basic info about enzymes and proteins, as it is too broad to cover in one or two posts. However, depending on what they told you and whether you accurately quoted it, it seems that you may be confusing things about protein and non-protein enzymes. Conjugated enzymes are still proteins, even if they are coupled to something else. Non-protein enzymes are required to be free of peptides or proteins. This include, for instance, ribozymes, which are made from nucleic acids. Again, read up first on basics of proteins, then go to protein structures (including primary, secondary and tertiary structures). Enzymes are a subclass of proteins, which you should read up after that. And only after you got a grasp of it I would go to enzyme kinetics and equlibrium reactions.

Sounds boring and the replay value is probably pretty lousy. May have good graphics, though.

Ok, so you are aware what you actually measure with this method (carboxylic groups). How many free carboxylic group does e.g. an intact dipeptide possess. How many if it is hydrolyzed?

Yes, the trendline would be called "calibration curve", but otherwise you got the principle.

Cancer is at least partially related to dysregulation of a number of cellular functions including cell proliferation and differentiation.

And you believe that if the media find people who elaborate on why the narrative of the news outlet is wrong they will actually show it? In contrast to, say, people like, Dr. Oz who can sprout popular nonsense completely unopposed? Face it, the media only shows what people like to hear. That is their main purpose and not to educate people. Unless you start your own news outlet you cannot do much against that.