dna - What conditions should I use for Gel Red staining?

I don't think you should need to vary the concentration of the GelRed. Mine came with instructions for the exact concentration and how to dilute. Optionally salt can be added, which I did, and it has worked for me.

I have only done this with Post-staining. 1-2.5% agarose with TAE. Used a 10,000x solution from Phenix research. The gelred stock and working solution needs to kept in the dark. I am assuming you have a transilluminator and (optional) filters to visual though. GelRed has similar excitation and emission [wavelength] properties as EtBr, so a standard transilluminator and filter should work without any changes. It will not be visible with the naked eye.

That being said there are a ton of things that can go wrong in casting the gel, eletrophoresis, staining, and visualization. If bands are visible but just blurry I was suspect there is more of a problem with either the gel creation or eletrophoresis though. Make sure agarose is completely clear and cooled down a little before pouring. Otherwise voltage is one of the main things I have noticed that can have significant affects on how sharp or blurry bands appear. You might try different % gels and different voltages. There are formulas that can help you estimate the best value for each of those.

evolution - Height and natural selection in humans?

@kate has what is probably the more correct answer for the observed pattern.

But as an experiment, I set up a basic simulation to approximate the conditions that you lay out:

1. Starting mean heights of 5'8" (172.72 cm) and 5'4" (162.56 cm) with standard deviations of 2.8" (7.112 cm). I used cm, because it's easier than dealing with inches.


2. Males will not mate with females that are taller than themselves.


3. Females will not mate with males more than 8" taller.


4. Males will not mate with females more then 8" shorter (follows from #3 above).

The problem that I quickly ran into was that, by truncating part of the normal distribution, the variance in height at each generation gradually decreased. After about 20 generations, the means weren't evolving because there was so little variation in height.

Human height is one of the most studied quantitative traits, going back over 100 years to some of the very first statisticians (Fisher, Galton). Height is a polygenic trait with very high heritability (h² = 0.8)¹. Genome-wide association studies have reported 54 genes involved in determination of human height².

Imagine that each of these 54 genes has just two alleles: a and b. a gives a +1 to height. b gives a -1 to height. So aa would be +2, ab or ba 0, and bb -2. The sum of all those alleles is correlated to height. So if all 54 were aa, then the height would be +108.

The problem comes in when people only mate with taller people. Over time, the proportion of b's will decrease, and the proportion of a's will increase, but only to a point. Once all the alleles are fixed at a, there won't be any room left. The genetic variation will be exhausted. Without the input of new alleles, height will cease to evolve.

1 Lettre, G. 2011. Recent progress in the study of the genetics of height. Hum Genet 129:465–472.

2 Visscher, PM. 2008. Sizing up human height variation. Nat Genet 40(5):489-90.

physiology - Dimensionless number for blood volume

Blood volume is not a dimensionless number - it's a volume. Historically we used to measure this in patients or volunteers by giving a large carbohydrate molecule like a starch that is not digestible or harmful to the body. Just like every other body fluid compartment volume (i.e. plasma, interstitial fluid, intracellular and extracellular) that we have, blood volume is estimated by intravenously injecting a known concentration of a particular compound. Once that compound equilibrates you take a blood sample and measure the compound's concentration again.

Initial Concentration * Initial Volume = Final Concentration * Final Volume

When you inject a known volume of a known concentration that only fills the "blood" component, and then you measure a final concentration - you can then solve for the "Final Volume" for "blood volume".

immunology - What cells would have the CD3 marker on them (other than T-cells)

As you know CD3 is

a protein complex and is composed of four distinct chains. In mammals, the complex contains a CD3γ chain, a CD3δ chain, and two CD3ε chains.

Natural Killer (NK) cells have been shown to express CD3 epsilon proteins, but not CD3 delta or gamma.

Also on a side note, there are several other non peripheral mononucleated cells besides Purkinje cells that can express CD3, but mostly in pathological circumstances. For instance Warthin-Finkeldey cells, also known as polykaryocyes, display CD3, although the origin of these cells is not certain (they might be multinucleated T-cells).

(PS: If you want more information on CD3, I found this site very interesting.)

neuroscience - Is the minicolumn the unit of the neocortex?

There are many arguments for what the unit of the neocortex is. "Columns" seem to be the standard, but what exactly those are is extremely contradictory between individuals, cortical regions, and species. Often times, when a column is referred to, it's actually a functional column without any anatomical borders (such as Hubel and Wiesels ocular dominance columns). Sometimes, the "column" is semi anatomical, such as the rat barrel cortex. Other times, these functional columns are confabulated into anatomical units, without any evidence for a border.

So my question is, could Mountcastle's "minicolumns" be the actual anatomical unit of the cortex? I've heard arguments that they are mere developmental relics. But they seem like the only reliable and consistent unit in the cortex.

physiology - Why is the human body able to repair a broken bone and not a heart muscle?

The heart does have stem cells in it, and there is cell turnover in the heart, of about 1% per year. Which is much slower than your skin, but not nothing. This allows your heart to grow during your life, and remodel itself slightly to become stronger/more efficient when you get in shape.

The heart can repair itself, when damaged it doesn't simply stay damaged. Unfortunately, the 'repair' leaves particularly useless scar tissue. After a heart attack, the dead muscle does repair itself, but very poorly. This 'scar' barely contracts, and isn't as strong as the heart wall around it.

This is mostly a function of the very specialized heart myocytes, and the evolutionary (relative) uselessness of being able to regenerate your heart after injury. In the wild, if your heart was injured, you were probably dead.

botany - Serological assays not detecting native proteins

Is there anyone out there who has done much work with serological assays? We have antiserum for a manufactured viral protein but no luck so far getting it to detect native protein (unless today's attempt worked which we will find out tomorrow).

What are any common problems or possible considerations that we should bear in mind when testing?