Hi Fernando, I would definately look at the enzymes from Kappa Biosystems as was mentioned in a earlier post - we have found these ezymes to be excellent. Their standard enzyme easily amplifies up to 15 kb.

Add the Grignard reagent derived from Me2CHCH2CH2Br (5 carbón atoms) to 3-Oxo-propionic acid methyl ester (hope the enolic form is not a severe problem).

Thank you dr jim

By the way, Richard, I note that Jon makes reference to a book that attempts to show how intelligence evolved intelligence, so the question clearly isn't his to answer.

Hello Gaber, I am trying to identify patches of brown adipose tissue in non fixed frozen cryostat sections. I have prepared Sudan Black saturated in propylene glycol. Counter staining will help to have an easier interpretation. Unfortunately, we don´t have any fast red around; so, just for a test I wanted to try with things available first. What is the role of Borax? Thanks, Cristina

Using Google you may found about 300 000 interesting articles on the question. See, please, “Comprehensive analysis of impact spherules supports theory of cosmic impact 12,800 years ago” (from May 21, 2013) Read more at: http://phys.org/news/2013-05-comprehensive-analysis-impact-spherules-theory.html#jCp

We also use RANKL from R&D (cat. #462-TEC-010/CF) at a concentration of 20ng/ml. You even get nice osteoclasts at 10ng/ml if you don't seed the cells to dense in the beginning, but to be safe i'll keep to the 20ng. I get osteoclasts after 3-5 days. It's important to check them regulary in this time frame because sometimes you'll have small osteoclasts in the morning and the day after the same cells are already apoptotic. And if there is no osteoclasts after 5 days they usually just proliferate and don't differentiate. That's the point where i thaw a new passage of cells. And you don't need M-CSF for RAW cells! However, for primary cells we use 25ng/ml of Cat# 416-ML-050/CF. PS: I had a minor crisis at the beginning of my PhD using other catalog numbers of R&D RANKL because the notion that you have to use a crosslinking antibody to this RANKL was well concealed in the small print >:C.

Hi, What you are experiencing is not uncommon in the plant world as the post above attest to. It is most likely your contaimination is due to the presence of polyphenols and quite probably polyphenol oxidases. The combination of the two lead to the production of reactiuve quinnones in the presence of oxygen. These reactiive compounds then interact with the DNA and RNA to both decrease yeilds in most commonnly used extraction methodologies as well as inhibiting downstream enzyme based manipulations such as PCR. All of the suggestions above will work to a greater or lesser degree. The addition of PVP and PVPP as well as increasing 2BME concentrations to between 5-10% works well. I have also found that removing phenol from the initial extraction (using only chloroform + isoamyl alchohol) also greatly enhances the yeild and purity of nucleic acids. Many of the commercial kits contain guanidinium based extraction reagents rather than CTAB and these may also cause you some issues. If you use the more traditional organic solvent extraction methodology I would strongly recomend using PhaseLock tubes from 5-prime - these act to provide a solid gel like interface between the organic and aqueous phases greatly enhancing your yeild and purity. THey are not expensive and the quality gains far outweigh the initial cost. A word of caution using column based clean-up kits - we have found that the quality of recovered nucleic acid is excellent the yeilds recovered are often quite low indicating quite significant losses. Also in my experience once you have oxidative processes at work in your extraction there is very little you can do to improve the quality of the resulting DNA/RNA....best to get the extraction right in the first place. Good Luck

A therapeutic strategy based on imaging is intriguing, provided that it has been formally validated. However, a couple of issues should be considered: the studies which tailored the duration of anticoagulant drugs based on the presence or absence of residual venous thrombus fail to avoid VTE recurrences in a low but not so neglibile fraction of patient fully recanalized. One more interesting issue is that related to the normalization of perfusion in a segmental pulmonary vessel after lung scan when the original intraluminal defect is partially reduced in size: is it a real phenomenon? should we consider it as a false negative recanalization of PE?

Heya, the inconsistency in expt may be due to reason that when you aspirate the supernatant you are loosing some cells. I have used this assay for suspension cell lines and it worked well. I seed 10^4 cells in 50 ul overnight and next day add 50ul of diluted drug, so that the total volume in well is 100 ul. After desired time point add 10 ul of MTT, incubate the plates for 3-4 hrs. Add 100 ul of solubilizing solution (10%SDS-0.01N HCl). Leave for overnight incubation. Next day read plate at 570 nm,. I got very reproducible results with this method. let me know if you face any other problem. Best wishes Loveena

Sounds reasonable. The foldX solution proposed above (or modeller) seems also good ones to me. Whatever makes your job simpler I would say.

Is useful the use of 0,01 % NaN3 to prevent bacterial and micotic growth, than to store the samples at least - 40°C

No, vice versum. Excessive illumination in not possible in UVVis spectroscopy. I think you are just getting 3A+ absorbance. Try dilution, recheck wavelength an make additional precautions if working in 180nm-210nm area

I've used Daniel's method before and it works perfectly :) good luck

Dear Matteo Glad to hear DMSO helped.This is the a first step to get rid of the proteins and possibly most of the biological contaminants. Now the question is what else is blocking you column. Could it be metal aggregates or is it fibrous material from dust/filters that is sitting in the fret or beginning of the column? If it is metals you can "reverse" wash your column with whatever solution/solvent that work best when you are preparing the metals for the nano-particles and that is compatible with the column. If it is the latter, you can try a bit risky last resort option - take the column of your system, remove the tubing and suspend just about 5 mm of the inlet part of the column in analytical quality filtered water or methanol and sonicate for 1-2 minutes, then put it back on you system in reverse and wash slowly with water or methanol of mixture. You can repeat this 2 to 3 times, but be aware that too much sonication can lead to matrix degradation and failure. Good luck! Regards Marina

maybe the link below can help you: http://www.quantiki.org/wiki/Quantum_Programming_Language#Quantum_Computing_Language

Providing access to great examples is a good place to start. There are some great tools out there that do fantastic things and use these to inspire your students. www.sliderocket.com is one tool which has some really great presentations that can be used as examples. Visual.ly also has some examples of developing presentations in innovative ways. Have a look at this great example: https://vimeo.com/66475816

It's been a while, but if you are still following this question i can only recommend NMR to you. It can provide quantitative data on bones (composition of the mineral or organic part, hydroxyapatite characterization, identification of amino acids in collagen, presence of citrate, presence of lipids, etc.) and only takes a few milligrams of bone.

Dear Tatini as many studies has been done on HPV in relation to OSCC lt they find no significant relation to smoking and alcohol consumption so Ithink of non sextual rout HPV -HR infection and because of latent nature of HPV infection make it difficalt to determine the real story behind its role ,but i still think the high % of HPV detection in OSCC could be non-sextual

Also physiologically the fish can be said to be sexually mature when there is a significant switch of energy allocation from somatic growth to development of gonads i.e. Sex cells and expression of sexual characteristics e.g. Colourations representative of the species.

I have used a number of tools including surveymonkey to zintelligence, surveygizmo and qualtrics. It depends on the skills of your survey designer and the complexity of the survey. My favourite survey program is qualtrics. You get 10 licenses for the same cost as one from survey gizmo but its drag and drop most of the way. The quality of the interface for the respondent is fantastic and you can do really nice visual questions that engage participants. The only annoying thing about it is that you cant email out a test link, but that's minor. It also has the ability to do really fancy skip logic and branching of respondent passage through a survey easily. I have only been able to do such complicated skip and filtering in Confirmit professional before, which is a really difficult tool to use.

From a slightly different perspective, The involvement of limbic - cortical regions of the brain is indicated. The uncoupling of the amygdala from emotions when BDNF is suppressed prevents new learning and possibly the continuation of disorders, that already pre-exist. The role of genes and polymorphisms such as 5-HTTPLR , COMT and others can be altered in their expression in the face of stressors, causing factors, such as high cortisol levels. I do hypothesize that fear which is the basis of anxiety, sadness which is the basis of depression may have differentiations in neurotransmission, and that should be there be higher levels of dysfunction in the limibic cortical region, that both anxiety and depression occur in conjunction, and reflect a continuum of dysfunction based on predispositions such as genetic, schemas, and social learning. I hope this is of interest.

Thanks for your answer. I have done redock implementing a water molecule into the binding cavity, but once I check the docking pose, it can be seen Autodock did not add any hydrogens. Therefore, any water-mediated hydrogen bonds were seen as expected. If the solution consists of allowing the water molecule be part of the protein receptor, how could I do it?