...used as a guide to point you toward a creative solution where everyone wins. What are the underlying...Importantly, as the third example shows, the ultimate resolution can involve ideas not related to the actual data... beneficial to both.  Coming up with creative solutions where everyone wins can initially feel more difficult... get unstuck and move closer to an innovative resolution. Creative negotiation can also decrease or entirely...with negotiating (check out more about conflict resolution). How to develop negotiation skills You can easily...being remembered as a person who tries to find solutions that benefit everyone. Even if I “lose” a negotiation...understand my position and to find a creative win-win solution. What does professional negotiation look like?...

...cells (9). PA-FPs have helped to revolutionize the field of super resolution microscopy. PA-FPs, especially...reversible type, are very well suited for super resolution imaging, where the power of the technique relies...reconstruct an image. The best PA-FPs used for super-resolution imaging usually have the following characteristics-high...wavelengths. Indeed, within the world of super resolution microscopy, PA-FPs have helped create several... improved over the conventional FPs for super-resolution imaging, they still do not emit the large number...being engineered. By combining PA-FPS with superresolution microscopy, scientists now have at their fingertips...V., et al. "Photoactivatable mCherry for high-resolution two-color fluorescence microscopy." Nature methods...

...typically load 3 dilutions onto our ddPCR plate. As the titer is unknown, each dilution should fall within...The process to titrate AAV by ddPCR begins with diluting the virus. It is important to note that the dynamic...:6 million to 1:25 million.  After making the dilutions, they are transferred to a new plate containing...reduction in the number of positive droplets as the dilutions increase. The ddPCR software uses the ratio of...Copies/uL V = Volume of virus in reaction mix D = Dilution factor of virus Most droplet readers have a few...separate area from where you prepare your sample dilutions. Aliquot all of your reagents into single-use ... pipetting too quickly when making your virus dilutions. This will reduce aerosols that could potentially...

... protect you and your institution, and almost all universities and institutions require them for material...protective of both the providing institution and the receiving institution. Therefore, many technology transfer...benefits for both the providing and receiving institutions and their respective researchers. Check out ...knowing your rights – and the rights of your institution – are protected while we handle all the paperwork... to you, or we can send it directly to your institution’s MTA coordinator. Organizations that have opted...

...-based directed evolution approach with some notable improvements. Directed evolution techniques mimic...you'll find in this post: Yeast-based protein evolution Fluorescent tools for mammalian cells Nanobody... Faster catalysis after yeast-based protein evolution By Joanne Kamens Mateo Sanchez and Alice Ting ...time modulation of the TEV cleavage and improve utility of the system for use with low affinity variants... new system has been demonstrated to work for evolution of other target proteins and might be useful for...screening systems at Addgene! Schematic of the evolution platform to generate TEV protease variants. Catalysis...transcription of the mCitrine gene. mCherry is constitutively expressed. Cells are then cultured and FACS...

...the location and distribution of synapses in the mouse brain. Kim et al is utilizing a mammalian GRASP...random mutagenesis in the soil bacterium Pseudomonas putida and demonstrate that they can successfully create...transport. pMitoTimer from Yan’s lab, which is constitutively expressed, reports on the balance of biogenesis...constructed using Golden GATEway cloning have been utilized to create recombination template vectors, to perform...chemogenetic technologies After several years of distributing his Designer Receptors Exclusively Activated...cloning vectors (Level 0, 1, or 2) which can be utilized in three successive assembly steps. Before beginning... (SSA) assay. This new Platinum Gate TALEN Kit utilizes a 4-module assembly system, which reduces the ...

...protein engineer currently working at Sanofi Pharmaceuticals in Cambridge, MA. When I joined Sanofi/Genzyme...including Activin A. A commercially developed therapeutic antibody that binds Activin A and interferes ...This is why we cannot stop searching for novel solutions to treat rare diseases like FOP until a cure is...fight against FOP in two ways. First, Addgene is distributing ACVR1 Plasmids from a variety of research labs...will be hosting the AJ’s Boston League team (Contributing many new heroes to the team in the process) ...protein engineer currently working at Sanofi Pharmaceuticals in Cambridge, MA. His work experience is in...in both vaccine subunit design and therapeutic biologic design (ADCs) along with pre-clinical analytical...

...Quality at Dimension Therapeutics, which was acquired by Ultragenyx Pharmaceutical in 2017. As one of the... the original employees at Dimension Therapeutics, she helped build and grow the gene therapy division...research funded,” she says. Baradaran met Addgene Executive Director Joanne Kamens several years ago through...Stegmeier is the Chief Scientific Officer at KSQ Therapeutics and is an Addgene depositor. KSQ uses a CRISPR-Cas9...Barcoding Library, a library that “enables the high-resolution tracking of more than 1 million cancer cells ...himself to finding “immunological and virological solutions to crises like those and ended up in the field...

...caspases, protease enzymes that cleave proteins. Executioner caspases are activated last in the cascade and...scientists have used genetically encoded fluorescent executioner caspase reporters. Several apoptosis reporters...Forster resonance energy transfer (FRET)-based executioner caspase reporters. In these reporters, the FRET... an improved apoptosis reporter, ZipGFP, which utilized the self-assembling split GFP (GFP1-10 and GFP11... of the linkers is a consensus caspase-3 (an executioner caspase) cleavage site (DEVD). Upon cleavage ...of sfCherry approximately 20 fold by directed evolution. They then optimized the length of both linkers...fold increase in fluorescence after cleavage by executioner caspases. Failure for cells to die or cells dying...

...dilute it, err on the side of caution when adding buffer back to the solution. Add a bit less buffer than...concentration to avoid over-diluting, and make sure to remeasure your titer after diluting to confirm. Luke...allow you to add many more viral particles to your solution while still using a volume that’s compatible with...surface of the filter membrane with the surrounding solution to ensure excess viral particles do not remain...remain adhered to the membrane. Be careful when diluting - if you end up concentrating your AAV above your...

...implement solutions for the stakeholders. Finally, software product managers test the solutions with the...cycle over again to refine the solution, or to come up with a new solution.  Now, you may be wondering how...the time, the problem to solve is clear, but the solution is more complicated. You’ll need to be creative...able to work with others to come up with a great solution.  How to get into software product management?...customer help tickets, and respond when I have solutions. We use these to help us know what to work on ...

...Nikolai Braun and Keira Havens, co-founders of Revolution Bioengineering. Read their previous blog post...the public in an open, responsible manner. At Revolution Bio, we are addressing this challenge directly...people can decide for themselves if they want this beautiful flower in their front yard or not. By developing...investigated on limited funding and time. While institutions that focus on creative problem solving like ... science, these tools sometimes have limited distribution. We hope that will change with Addgene’s expansion...project. We’ve shared some of the concerns that Revolution Bio had to address as we developed the color-... in the United States Air Force. She left active duty to pursue a degree in a synthetic biology laboratory...

...description of the RFA and funding institution. Emphasizing how your solution fits into the mission of the ...funding from a variety of government and academic institutions. New graduate students may assume that their...advisor may choose to allot some if not all grant duties to trusted students. For example, if the proposal...the mission of the application and the funding institution. Moreover, avoidance of ‘to be’ verbs creates... of the statement, use it to confirm that your solution is the best approach to the research challenge...the intended protocol, and offers alternative solutions to demonstrate thorough thought planning on the...

...These events have also started progress towards solutions. The 2015 Boston FoR meeting was designed to explore...further data is necessary to determine the best solutions. The event also culminated in a joint American...students and postdocs broadly – and thus PI’s and institutions in turn. The first is the passing and upcoming...the NIH recommended pay scale – used by many institutions to set postdoc wages. The second event of note...simply being able to categorize all postdocs at institutions, will be challenging. At the same time, there...level, FoR is currently compiling a database of institutional responses to the FLSA ruling. Similarly, from... and graduate students, and started to define solutions but this year, together with Academics for the...

...expression following transduction with serial dilutions of a viral prep. This method is particularly useful...Fluoresence Titering Protocol Another popular approach utilizes the antibiotic resistance genes carried on transfer...plasmids. Target cells are transduced with serial dilutions of a viral prep, treated with the antibiotic, ...intensive. Target cells are transduced with serial dilutions of a viral prep, genomic DNA isolated, and primers...months but should be considered in labs that will routinely be producing virus or require precise titers. ...their benefits and drawbacks. Labs that do not routinely produce virus may want to choose more straightforward...colony-forming unit assays, whereas labs that routinely produce virus may want to consider the more accurate...

... their PhD and postdoctoral training. Addgene executive director Joanne Kamens has written a 5-part blog...pursue leadership opportunities at the local, institutional, regional, and national levels. These skills... But how does one gain such experience? Your institution may offer occasional workshops on leadership ...Harvard Medical School and then Boston University (institutions where I’ve worked as a postdoc). I soon realized...that many of the issues faced by postdocs and institutions are not local but are national issues. I applied...many changes to the  Board of Directors and the Executive Director changed twice. I had to learn how to ...to the Board of Directors and liaise with the Executive Director. As members changed, I had to learn how...

...that is shared across all CRISPR types; the fast evolution of these adaptive bacterial and archaeal immune... in archaea, though Class 1 remains largely underutilized compared to Class 2. Class 1 Despite their incredible...., & Koonin, E. V. (2014). Classification and evolution of type II CRISPR-Cas systems. Nucleic Acids Research... E. V., & Makarova, K. S. (2019). Origins and evolution of CRISPR-Cas systems. Philosophical Transactions...Zhang, F. (2017). Diversity, classification and evolution of CRISPR-Cas systems. Current Opinion in Microbiology... J., . . . Koonin, E. V. (2015b). An updated evolutionary classification of CRISPR–Cas systems. Nature... Yakunin, A. F., . . . Koonin, E. V. (2019). Evolutionary classification of CRISPR–Cas systems: a burst...

...spectrum and emission spectrum. Fluorescent dyes utilize these fluorophores by combining them with a molecular...These variations directly impact the way you can utilize these dyes. The most-used fluorescent dye is fluorescein... between multiple dyes during analysis. Image resolution is impacted by several factors. Firstly, brighter...have been specifically designed for use in super-resolution microscopy techniques, such as Stimulated Emission...These dyes have specific properties that enable resolution beyond the diffraction limit of light. Dyes like...properties that position themselves for super-resolution usages, such as superior brightness, photostability...brighter and more stable fluorescence for super-resolution applications. Other current advancements include...

... sticking pipettes into solution bottles directly, rather decant the solutions into disposable sterile... up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine. Twice...that you also have the power to take the added precautions necessary to keep your cells from becoming contaminated...things more manageable, but it also keeps stock solutions from getting contaminated. Make sure cells don't...So if you, for example, split cells on a Monday diluting them 1:10, you should be able to split on the ...

... positives [2, 4]. Solutions to the polymer problem There are some simple solutions available for researchers...GFP to study the protein's behavior in cells, in solution or something in between, you are using a tag with...ruin your day (and experiment) are listed below. Solutions to avoid these all too common issues follow. ...with all of these problems. The simplest and best solution is to use truly monomeric FPs. EGFP and other ...they are designated mEGFP, mVenus, etc. A word of caution is needed because FPs not derived from GFP, i.e...depend on protein concentrations. An imperfect solution is to restrict imaging analysis to cells expressing...