Pardon my very late entry into the discussion. My pink chicken was distracted by other concerns. While I suppose there is some intellectual fascination in trying to establish a bright line between the bizarre and non-bizarre, in clinical practice the difference is usually obvious. I had a patient who believed that beings from Titan who were engaged in a civil war had switched venues to his body in order to spare their world from damage. Bizarre. A colleague had a college student who complained that the CIA was poisoning his toothpaste. Non-bizarre, by the usual rule of thumb. On the borderline is the elderly gentleman that I saw who claimed that all of his neighbors had been replaced by Cuban lookalikes. (Why Cuban? I have no idea. This was central Illinois!) I would argue that deciding whether this final example is bizarre or non-bizarre is of no demonstrable diagnostic value. So, why bother?

Just as the "physical electron" is an infraparticle and does not correspond to the non-interacting "elementary particle" or field excitation which appears in perturbative quantum field theory, quark fields just serve as coordinatizations of the underlying theory which has asymptotic states as protons (if stable) etc. An electron is dressed by the electromagnetic field, so is it elementary? What a quark is depends on the way you look at it. From a very naive point of view, one could argue that if there were subquarks, they would be confined to a very small region, implying a cancellation of high momentum and kinetic energy versus binding energy, since otherwise they would have a large mass. This fine tuning problem then could be related to a symmetry of the "subquark" theory. Such considerations, however, are dubious at best.

Absolutely yes!

Thank you all for the answers. I tried washing the column first with buffer that had 0.5M Imidazole. This turned the column very blue. I then washed it off and re-equilibrated it. also, I did not put any DTT in the binding or elution buffer, just in the lysis buffer. And it worked..!! I have some protein now. I appreciate all your suggestions.

I've successfully used Sigma AccuTaq LA to make 30kb products from E. coli genomic DNA. It's a mixture of Taq and a proof reading polymerase (as Joachim suggested).

1. You say the drug to be insoluble in DMSO and soluble in media, wondering why not in water which makes up the media. 2. The media (animal cell culture) in general cannot be stored for more than 2 weeks, depending on the stability of its components. Some turn colour showing the change in pH. Such change in conditions like pH can affect your drug properties. I would not recommend storage in such a medium rather than a well characterized solution or chemically neutral/stable one like water. 3. There is an inherent problem here in your condition. Media is meant to be for growth of organisms. In case you have sterilised (filter) your media the drug can be there for some time (few days).

Try OPENSTAT open ware/ free download available

I already am aware of the definition. All I am asking is whether there are any examples of a system where a mesopredator is larger than the apex given that a meso and apex predator are defined by their trophic level rather than body size, despite the usual relation of mesopredators being smaller.

I think SurveyGizmo is the best online tool out there. You get way more with the free version than any other option. I've used it for academic work, and it was easy to design, edit, implement, and analyze my survey. I will never use surveymonkey again.

Personally, the best way to rapidly quench any samples for any further mass spec analysis is use a dry ice-ethanol slurry. Then dip a tube containing you samples in that slurry and mix with a thermometer in order to avoid freezing of your cells at the bottom of the tube and check the temperature. When it reaches 8-10C, you cells have already reached 4C and metabolism is quenched. Then, you immediately transfer your tube on ice and carry on your metabolite extraction (at 4C the whole way). The quenching process should not take more than 2-3min. (Same method as what Juliano de Toledo mentioned above for Leischmania). This should answer quenching issues with any eukaryotic cells.

Thank you guys for you suggestions. I just wonder if i do multiplexing in one PCR tube then how to run gel for the products? I mean, all gene products are in same tube then How can i compare with housekeeping gene? can anyone give any picture?

Thank you Andjelka, I have downloaded it and will give it my best attention.

Thanks for the clarification, David.

That i knew as for a vertex cover it has to cover all the edges

K-means is a simple method that is often used to partition (cluster) the data in an unsupervised way. The centers move around over many iterations until they converge to a local optimum. Different initializations can lead to different optima (depending on the basins of attraction) - so the clustering is usually re-run several times and the best result is taken. Alternatively, more care can be taken while initializing the centroids - look for example at the K-means++ K-means can be easily kernelized and Kernel K-means allows for detecting clusters that are not (hyper)spherical in shape. Spatial indexing can be used to speed up the search for the closest centroid, by pruning out the obviously distant ones (which is not crucial for small K-s, but almost necessary for larger ones - in my implementation it speeds up the clustering by a factor 5-10, depending on the data). Also, the K-means search can be a bit stochastic in order to avoid converging to local optima and guide the final configuration to a global optimum. We have recently proposed one such method, the Global Hubness-Proportional K-Means (GHPKM) that performs rather well on high-dimensional data. The details can be found in the journal paper here: http://www.computer.org/csdl/trans/tk/preprint/06427743-abs.html

I think that problems are not in the methods or sophisticated protocol of extraction DNA. Maybe the problems are in the condition or standarization of this methods in your lab. I am refering that all that your are used in the purification. You will need establish a control of quality that garnanted that all is ready for used in one purification. And after I recommend checking a simple and easy protocol for extraction DNA to use in AFLP.

A couple of suggestions to the many already proffered should the problem continue AND should the material in question be viable (many seemingly sound seeds aren't): - Even if the Cæsalpinia sp in question is not a pyrophyte, the heat in rotting manure - sometimes used with slow-germinating oil and other palm seeds - may accelerate germination. Placement in warm to hot decomposing compost for days (¿or weeks?) may have a similarly effect. - Some seeds putatively require passage through a herbivore's gut. Might that be the case here? I hope you will tell us in due course what strategy worked best.

enhance dilution of the analyte until 10-5 Molar or use another deuterated solvent as dimethylsulfoxide sometimes its works

I found the following resource quite helpful. I had similar questions as yours.

Well, if the two brains are not from the same subject, then you can't expect the ventricles to align perfectly since their anatomy will be locally different. If they did align perfectly, then one of those datasets would have been deformed so much that you may not be able to infer an accurate location for the differences you might observe. I believe that this problem can only be resolved at the group level (i.e. comparing two groups rather than two individuals) if your two brains are from two different subjects.

Which protocol are you using? Is it published?

Dear Valesac, Euphorbia may be pretty tricky due to the milk-like terrible substances that some of these plants have. I recommend to use a technique that relies in a first step on rapid degradation of everything that attacks RNA and on density as major parameter for separating RNA from all of the rest. This should probably work. I have used the technique below over several decades for Zea mays (sometimes difficult due to high starch content, depending on the developmental stage of cobs and kernels) and for many different fungi, plant pathogens and others. Now the citation from a previous answer that I recently gave to a colleague, decribing the procedure. "You depend on complete and fast denaturation of proteins during extraction. The best you can do is grinding in liquid nitrogen followed by denaturing in detergent/guanidinium thiocyanate (very good) or urea (good but not quite as good) and mercaptoethanol. The procedure provides you with high molecular weight RNA from any biological material. For separating RNA from DNA, protein and lipid it is very successful and easy to centrifuge the extract through a small CsCl cushion in a swinging bucket ultracentrifuge rotor, which is essentially no work at all. You end up with high yields of very pure and undegraded RNA. For experimental details see: Translation of Zea mays Endosperm Sucrose-Synthase mRNA in vitro. Johannes WÖSTEMEYER, Ute BEHRENS, Armin MERCKELBACH, Markus MÜLLER, ,Peter STARLINGER. Eur J. Biochem. 114, 39-44 (1981). (The urea procedure) or Fungal Genetics and Biology 42 (2005) 804–812 4-Diihydromethyltrisporate dehydrogenase, an enzyme of the sex hormone pathway in Mucor mucedo, is constitutively transcribed but its activity is differently regulated in (+) and (-) mating types Christine Schimek, Annett Petzold, Kornelia Schultze, Jana Wetzel, Frank Wolschendorf, Anke Burmester, Johannes Wöstemeyer (the guanidinium thiocynate procedure)." It works, and you don't need expensive kits with more or less unknown compositions. - Good luck!

Here is another approach for totally qualitative data - where quantification is not possible. Plot the data points - Use ArcGIS or QGIS. Create theissen polygons for the location points. Now based on those points - assign soil attribute to each individual polygons as it appears for that location within the polygon. By these you will be able to generate a crude soil type dsitribution map. Now use all other background information available like elevation, relief, slope, drainage, depth of weathering, geomorphology, geology etc to finally derive at a pertinent result. Fine tune the same with the available data and records for that area. Hope that helps

Given your ordered categorical outcome variable, running t-tests will most likely yield biased estimates due to the normality assumptions being violated. Normally, I would fit an ordered categorical logistic regression model and include the exposure using dummy variables but in this case I'm worried that you would lack the statistical power to run this model due to your very small sample size. Perhaps a more descriptive approach would be better in this specific case.

I do not think you can elute the biotinylated DNA from streptavidin using just biotin and non-denaturing conditions. However, you can do that if you use mono-avidin beads (Pierce, Adar Biotech). Biotin binds to mono-avidin 10 to the 7th-fold less than to streptavidin tetramers. See: http://www.piercenet.com/browse.cfm?fldID=01021104 One way to get the DNA-biotin off of the streptavidin "may" be to use a protease to degrade the streptavidin, you could try protease K along with 2% SDS (but SDS is not required) at 24-37oC, and then add 2 mM biotin. Once the Pro K has degraded the streptavidin biotin should be able to compete off the DNA-biotin from the monomeric streptavidin. However, the problem may be that the streptavidin is no longer attached to the beads, so you will end up with DNA-biotin with free biotin, small peptides, and of course Pro K.

In your case that would be two genes from duplicated chromosomes (homeologs). Any gene on one chromosome in the A set should have a counterpart in the B set of chromosome, unless the gene had been lost.

I really do not know how fast Dexamethasone or other drugs may Need to reach their full pharmacological potential, but I could Imagine that even a satisfactory blood level of dexamethasone after a few minutes may not reflect blockig of all pathways in all cells and tissues. I also could Imagine that just your two windows of 5 and 10 minutes could results in different results. I could be wrong but would Check how fast it gest into the cells of interesting. Blood Levels may be Important for drug-Monitoring AFTER the workung level is reached.

In response to Heidi Kocalis: Dr. Lustig does not claim Fructose alone causes obesity. He clearly discusses its relationship to metabolic dysfunction, even in the absence of obesity.