...ligation mix in different bacterial strains (stbl2, NEB stable, etc.), and/or use a different temperature ...want to assemble together in a combination of different ways according to your cloning strategy: Restriction...buffers and temperature are compatible between the different enzymes. Always check the fragments size of your... There are multiple ways you can assemble the different parts of a plasmid based on the cloning strategy...attention in the areas of ligation between the different fragments If you don’t get any positive clone...might want to reconsider your approach, and try a different cloning strategy. Once you've identified potential...Nature, and other journals.   Paolo Colombi is currently the product development scientist at Addgene. ...

...section, teams are assigned different groups of three genes selected from different parts of the Met pathway...first select a multi-gene pathway to study and a reference organism where deletion strains are available....Students next transform deletion strains from the reference organisms with the overexpression plasmids. Finally...species has a similar function to the gene in the reference species. We have been using yeast models (3, 4...yeast, Saccharomyces. cerevisiae, makes a great reference organism for the project because it is non-pathogenic.... Over time, the two genomes have become very different due to a whole genome duplication and extensive...cloning to insert the coding sequences for ten different MET/Met genes (Figure 1) from the two yeast species...

...engage broadly with diverse communities, to raise awareness about where science is and where it is going, ...Personal Genetics Education Project (pgEd), where I currently work, strives to broaden the conversations about...Harvard Medical School, has a mission of raising awareness and starting conversations about both the benefits...and epigenetics. So, I sought out a number of different opportunities to broaden my training beyond the...major part of our everyday work is to liaise with different organizations and individuals, to identify ways...collaborating, and to organize events that bring different voices together to talk about genetics. In addition... such as scientific, educational or industry conferences, as well as schools, community events, churches...

...of the Canton Nucleic Acids Forum (CNAF) 2015 conference invited me to attend and speak about Addgene’...attended by over 300 Chinese scientists eager to hear current findings in nucleic acid science. I was delighted...Thoughts on Guangzhou and the Science City The conference was located in the modern, relatively new “Science...national science and technology park, it enjoys preferential governmental policies for a high-tech development... which I could identify. There were dozens of different mushroom varieties alone. At least I could tell...requires an interdisciplinary team.  Day two of the conference was kicked off by Nobel Laureate (and Addgene...circRNAs do yet or how they do it. Of course, No conference would be complete without CRISPR Michael Terns...

...to learn how to adapt to different situations and, in particular, to different styles of leadership. As...must accommodate many different types of people in order to harness their strengths and work effectively...to learn more. Having met many people at that conference, I later became involved with the NPA beyond ...within the committee, I became Vice-Chair when the current Vice-Chair stepped down. When the Chair then stepped... the Committee which I then led for 5 years.  Different leadership styles Being an effective leader requires...requires you to understand how to work with many different types of people both above and below you. During...relationships with these individuals who often had different styles from their predecessors. Changes were implemented...

... or looking to improve your current methods, understanding the differences can help you make an informed... Resources References Shaw, G., Morse, S., Ararat, M., & Graham, F. L. (2002). Preferential transformation...make the virus. Viruses can be produced in many different cell types, spanning both prokaryotic and eukaryotic...However, some viruses can be produced in multiple different cell types, including plants, mammals, and insects... transfer plasmid. The transfer plasmids have different packaging capacities depending on the virus. AAV... which system to use will come down to which differences are most important to you — virus type, cost,...this post has provided some insight on these differences in viral production and will help you decide ...

...first successful attempt to render fixed anatomical preparations transparent was done by Walter Spalteholtz... Pubmed. J. Sharpe, U. Ahlgren, P. Perry, Hill B, Ross A, J. Hecksher-Sorensen, R. Baldock,  D. Davidson...microscopy coupled with techniques that make tissues transparent are enabling microscopes to visualize the cellular...spinal tract, CST, spinal cord and cranial nerve afferents labelled with a AAV8 expressing eGFP under the...sample and captured the reflected light using a different objective lens oriented perpendicular to the plane...of clear specimens and for in vivo imaging of transparent organisms. Advances in tissue clearing Similar...Find plasmids from the Tsoulfas lab here.     References: H. Siedentopf, and R,. Zsigmondy.  (1902) Ann...

... This preferential tropism for different brain regions is largely due the ability of different serotypes...tissue? What is commonly known is that different serotypes have different tropisms but, as you can see, AAVs...there are marked differences among serotypes in their efficiency at transducing different brain regions....serotypes to infect different cell types. In the CNS, it has been shown that there is variable transduction...skeletal muscles, and the central nervous system. Currently, AAVs are being used to deliver CRISPR/Cas9 but...transduction efficiency, and safety profiles should strengthen its appeal for the scientific and medical communities...movement. Follow her on twitter @katrinaboards.   References 1. Russell, David W. "AAV vectors, insertional...

...presenting your research at conferences such as SACNAS and ABRCMS. These conferences have a lot of programming...her on Twitter @EleanorWang.     References and resources References Canning EA, LaCosse J, Kroeper KM...new therapies can be developed and applied. The current pandemic serves as a striking example of the applicability... is a supportive environment—it makes all the difference in a lab experience. “Mentorship” was a bit of...interested in doing research and attending research conferences. The Barry Goldwater scholarship is also a national...graduate programs, recruiters will attend these conferences to answer questions and offer application fee...explore! Addgene has an entire page dedicated to different career options from science policy to science ...

...across multiple teams or to stay abreast of current trends in the literature, sampling a breadth of work...One thing many scientists enjoy is discussion of current new and exciting literature. As I transitioned ... had concerns that I may not have time to stay current or enjoy that discussion. Luckily, one needn't ...science, researchers of all varieties must stay current with literature and emerging technologies to remain...meeting amongst scientists that aims to help stay current on new and emerging work by collaboratively discussing...the meeting settles in and people get the time recurrently carved out. This notice typically should include...the same figure by figure organization with a different person responsible for each figure. Others still...

...immunotherapy than ever before. Although different types of cancer have different challenges and obstacles to overcome...However, there are many different forms of immunotherapy that work in different ways. For example, a new...learned much about the causes of cancer and the different avenues that can be used to treat it. For those...latency period, and rarity of mesothelioma make it different from many other types of cancer. Mesothelioma ...literally thousands of years to Ancient Greece. Current mesothelioma treatment As with many forms of cancer...catch the disease earlier. New techniques are currently being tested in clinical trials to detect mesothelioma...mesothelioma, such as in the case of Paul Zygielbaum. A different approach to immunotherapy is bevacizumab (Avastin...

...in the U.S., whose current salaries are below the new exemption level (the current average postdoc salary...exactly how postdoctoral salaries are changing. Transparent salary information for postdoctoral positions...data) and therefore the numbers of postdocs at different points on this salary range. We have been able...are doing, however, may face greater challenges. Current salary information that we have been able to find... with relative certainty that just over 40% of current postdocs will have their salaries raised to the...already have salaries at or above $50,000. Boston Children’s Hospital is the only part of the Harvard group...laws will have on the postdoc population. In the current funding climate combined with the lack of a sustainable...

...strains of mice. This suggests that a small genetic difference between these two mice might drive their divergent...took a broad approach and searched for genetic difference between the 36 mouse strains that are part of...permissive phenotype, but only one, Ly6a, showed differential protein and RNA expression between the brains...level of CNS PHP.B transduction as their C57BL/6J parent. The second generation (F2) of outbred mice also... if PHP.B required the same receptors that its parental AAV9 relied on for transducing cells: galactose...These results suggested that PHP.B, unlike its parental AAV9, does not require galactose and AAVR to transduce...are not reproduced when the vector is used in different mouse strains. The in vivo selection for PHP.B...

... new prime editors, PE6a–g, that overcome some current prime limitations and enable impressive editing...optimized for dual-AAV systems. These variants use different RT enzymes and performed large edits with similar...similar or better efficiency than PEmax, the current state-of-the-art. PE6b was very efficient for short...types. DIRECTED is compatible with fusogens from different viral families and can be used to target multiple... enzymes, TurboID and APEX2, are localized to different cellular compartments. They showed APEX2 can use...uses sequential labeling by TurboID and APEX2 at different cellular locations. C) Enrichment of dual-labeled...stably or inducibly express tagged iPAK4 from different promoters, specific time points appear as fluorescent...

...these tools can be to use. AAVs, which can target different cell types depending on their serotype, are particularly...93,491 viral vector preps with researchers in 45 different countries — and as of 2024, we’ve distributed ...the vectors in Addgene’s catalog are offered in different serotypes. In fact, the 879 viral vectors in our... CRISPR, where we dove fairly deeply into the different types of tools scientists were using/depositing... is less amenable to such analysis, due to its inherent bias: Addgene uses a prioritization process to...service. But why are their request patterns so different? It could be a matter of timing, as jGCaMP7 was...from requestors and depositors, and identifying trends in our own repository data, giving us significant...

...more scientists in our leadership, and raising awareness regarding the pitfalls of accepting alternative...outside the scientific community. Vanessa Sung is currently a PhD student studying breast cancer at the Goodman...Over the last few years, there's been a disturbing trend in the number of high profile people denying science...climate change is a hoax. This has culminated in the current President holding and campaigning on many views...Candidate studying Nuclear Science and Engineering at Rensselaer Polytechnic Institute. He also studies energy...that I expected. Jennifer Stancill, Ph.D., is currently a post-doctoral fellow at Vanderbilt University...fingers of skepticism to defend an agenda that halts current data collection, destroys existing data, and guts...

...can't distinguish the different proteins from each other. To distinguish different target proteins in your...target. By using a mix of different antibody-fluorophore combinations for different markers of interest, you...fluorophore. However, these aren't strict rules and in practice the differences in brightness levels are ... fluorophore's brightness refers to the signal strength of the emitted photons detected by the cytometer...a global maximum at around 560 nm, leading to different levels of brightness, depending on the laser light's...floodlight. For fluorophore brightness levels, these differences are much more subtle (think of brightness levels... Even though the two compounds are completely different in nature (eGFP being a protein and FITC an organic...

...numerous variants for the different types of opsins available and their differences and uses have been discussed... the different variants [1, 2, 4]. And, of course, the best way to learn about the many different opsin...particular neurotransmitter release or post-synaptic current. Fortunately, if your lab focuses primarily on ...about this in detail later). But what are the different steps in an optogenetics experiment and what are...Biology from the University of Michigan and he is currently a Postdoctoral Fellow in the lab of Mary Jeanne... or follow him on Twitter @derekpsimonphd.    References 1. Tye KM, Deisseroth K. Optogenetic investigation... for analyzing the neural circuits of behavior. Trends in cognitive sciences. 2011;15(12):592-600. doi...

... science) they come from many different places supporting many different positions. It would be detrimental...cycle begins again. No part of this process is inherently political so why is there a march in Washington... (scientists who hold a doctorate degree, are currently working full-time, and have received a research... and everyone is welcome. Why? There are many different things scientists and non-scientists alike are...not necessarily marching for every cause. The references provided also only provide a snapshot into the...microbial interactions, and science education.       References March for Science Website Locke, John, and Peter...Cambridge university press, 1988. AAS: Historical Trends in Federal R&D NJ.com: Rush Holt: Lawmakers need...

...deposits to Addgene.  Insomuch as different journals represent different research areas, target readers,... this story had us wondering: is there a larger trend here? Do papers associated with Addgene deposits... would also be great to know how much of that difference in citations is caused by depositing a plasmid...wouldn’t be causal. To control for a number of differences between those papers with Addgene plasmids and..., so our estimates now take into account any differences in citations due to the journal of publication... the same journals! So, is all this remaining difference caused by depositing the related plasmids with...it is still suggestive that we see such a big difference in citations after controlling for journal. And...