...the gel and add gel components to polymerize. Step 3: digest the sample to allow for even expansion.  ...:832–863. https://doi.org/10.1038/s41596-018-0117-3 Wassie AT, Zhao Y, Boyden ES (2019) Expansion microscopy...applications and there’s just not much to be done about that.  Second, expansion introduces distortions....

...been trafficked between locations). Figure 3: A) LOV-Turbo becomes active when illuminated with... h, and PMA activation induces cFos::GFP-iPAK4 at 3 or 6 h. Scale bar 5 µm. Images adapted from Lin et...section of the Addgene antibody page, and learn more about the whole-brain reference atlas efforts, including...

... from and how the idea got off the ground Chapter 3 - Addgene as a leader in standardized tech transfer...repository’s future. If you’ve ever been curious about what it takes to start your own nonprofit, this ...Harvard University, where she studied diabetes and metabolism. Melina has worked at Genetics Institute in Cambridge...

...not immunogenic, a necessity for in vivo studies; (3) is compatible with single gene KO and pooled screening...faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment that exists in ...LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School and is interested in...

...the impact of research" (2013) Scientific Reports 3, Article number: 1649 doi:10.1038/srep01649     ...Of course, all of you toiling away in laboratories this holiday season want the work you are doing to... has to do with online accessibility. It is now about as easy to find a paper in a lesser-known journal...

...Improved Vectors for Gene Delivery. Curr Gene Ther 5(3), 299–310. PubMed PMID: 15975007. PubMed Central PMCID...Concepts in the pathogenesis of rabies. Future Virol 3(5), 481–490. PubMed PMID: 19578477. PubMed Central...viral vectors is serotype-dependent. Gene Ther 20(3), 348–352. PubMed PMID: 22418061. PubMed Central PMCID...to its retrograde axonal transport. J Cell Biol 82(3), 798–810. PubMed PMID: 92475. Taymans, J. M., Vandenberghe...usually severely impaired, and only few reports exist about dye molecules that are transported from one neuron...

...up the experiment. The first way, it will take you 3 days to complete the entire experiment, but you’ll...be quite difficult. In my experience, planning has 3 components: A mechanism for blocking off time for...post useful. After a bit of research and thinking about my own processes, I’ve broken down time management...task, you’re actively prioritizing. In my anecdote above, I prioritized splitting cells over meeting up with...was too late. I didn’t spend enough time thinking about the possibilities and how to prioritize them. When...month, or even the year, you should be thinking about two main things: How useful the task is (its utility...plasmids to send to Addgene). Following my advice from above, you should probably take the second option. Even...

...understand your proposal to give you proper consideration. 3. Present an easy to follow seminar See item 2. Attendees...? How will your approach resolve this problem? (2-3 sentences) What will be the focus of your first... potential colleagues study. Think about how you might collaborate or, at the absolute minimum, can appreciate...the first sentence or two what your research is about and what you think you can accomplish. No matter...your seminar accessible. Let the experts ask you about the details and then you can display your deep knowledge...expert's question than to leave everyone confused about your research because you waded too deep into the...simple in principle The first question I usually get about chalk talks is, what is a Chalk Talk? It is a 60...

...limitations. 3. Always provide value to others This is a non-profit organization. Therefore, collaboration and ... of how much you can expect to use in the future. 3. Identify the skills needed to use these tools Outline...a successful negotiation and collaboration. We’d love to hear more about your experiences and opinions...can get away with using computer modeling and collaborations rather than using animals, then that’s a cost...associated with old equipment, like service repairs, labour costs, and loss of time, can exceed the cost of...Everything is negotiable. Try asking for discounts, collaborations, access to internal award applications, etc...Happy Sciencing, Damien   Damien Wilpitz is a laboratory research management consultant and coach in Boston...

...back to the drawing board.  Cycle 3: A Return to HQ Figure 3: Even Blugene is practicing social... Production Specialist, I’m constantly thinking about ways to improve our productions to deliver the best...four months after I started with Addgene, I was about to have a check-in with my boss Jennifer. I had ... would be a couple months. Jennifer and I spent about an hour that morning scrambling. Everything we had...someone to be comfortable on camera while thinking about things like lighting, camera angle, and shot composition... allowing people to sit without having to worry about getting in close to mic them up. Monitor: I then...first video in our Intro to the Lab Bench series about PPE! We made this with a combination of in-lab filming...

...the world — and we really do mean all over (Figure 3). We continue to add new posts every week, and we’...beautifully designed and easy-to-reference eBooks. Figure 3: Global readership of the Addgene blog in 2023. Views...a less tangible form: knowledge and information about the materials in our repository. We started to increase...by making it easier to find materials and learn about them. As early as 2007, we began grouping similar...professors, stopping by to say, “Everything I know about virus, I learned from Addgene,” or “I learned to...seeing our resources shared via social media, on collaborators’ websites, or as a recommendation in a Reddit...updating and expanding our website, we’ve learned about the importance of smaller, systemic changes, like...

...experimental design and gel quantification. Figure 3: In a perfect world, protein concentration in your...versus the amount of protein loaded on the gel (n=3). Data is presented as mean ± SEM. Adapted from Analytical... this chain (blue) to infer or calculate things about earlier ones. In practice, multiple sources of error...condition 2.” By eye, the loading control appears about the same between the two conditions. But it isn’.... However, it throws both assumptions discussed above into question. Assumption #1: The loading control...searching the literature to see what is already known about your housekeeping protein in your context. If you...step from the POI blot, you don’t need to worry about the linear detection ranges of multiple targets....

... originally written by Melina Fan and updated Nov 3, 2022 by Susanna Stroik. You’ve designed your gRNA...PCR and a restriction enzyme digest.   Fig. 3: Restriction enzyme screening for resistivity to digestion... your edit, it’s important to know a few things about your target cells and the desired edits. First, ...et al., NAR). It works just like TIDE (outlined above), except that it uses a 3rd sequencing trace file...great option, especially if the options outlined above aren’t realistic. Check Out Our Post on Sequencing...higher probability locations. If you’re worried about off-target effects, you can consider using a version...SpCas9-HF1, eSpCas9(1.1), or HypaCas9. Read more about Cas9 options in our blog post here. Dealing with...

...conservative estimate. As a graduate student, I drank 3-4 cups of coffee a day, and I had colleagues who were...learn about all the amazing research being done with plasmids. I heard an impressive talk about the use...pulls off such an inspiring, thought-provoking, collaborative event – all while keeping the party going! At...scientists we've spoken with are excited and passionate about making such tools open-access. Though it is time...

... why they are the most commonly requested (Figure 3). Over 400 biosensors were distributed in 2023 alone... viral vectors that deliver GFP.   Figure 3: A) Cumulative AAV preps available at Addgene from...all, a delivery method. I would be remiss to talk about them without looking at what they’re delivering .... But in the years since, we’ve learned so much about what it takes to make these viral vectors not only...., 2016), video, educational resources, and collaborations, and will continue to do so, in the hopes that...leave a comment; or take our AAV survey. Learn more about GCaMPs and other tools developed through the GENIE...

... them. For intracellular targets, we usually test 3 different fix/perms (methanol, PFA-saponin, PFA-triton.... The approach we suggest is summarized in Figure 3. The principle is to confirm the antibody can detect...-use of an individual experiment.   Figure 3: Our recommendations for an approach that may be used...leukocytes. The file shared also contains information about the labeling pattern of several cell lines using...Laflamme, C., Virk, H., NeuroSGC/YCharOS/EDDU Collaborative Group, & ABIF Consortium. (2024). A guide to...

...startups, and real applications of SynBio. Chapter 3: Synthetic biology as the engineering of biology at...synthetic yeast chromosome. Listen to learn more about Dr. Ellis, synthetic biology, synthetic chromosomes...Additional Resources from the Addgene Blog Learn about the Synthetic Biology Education Nonprofit BioBuilder...BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup in Our Interview with...

...negative controls for genome-wide CRISPR screens; and 3) safe-targeting guides, which target genomic sites...PMID: 25513782. Pubmed Central PMCID: PMC4320685. 3. Fu, Yanfang, et al. "Improving CRISPR-Cas nuclease...following unanswered questions about CRISPR screen design. Unanswered questions about genome-wide CRISPR/Cas9...Morgens et al drew some generalizable conclusions about off-target activity, demonstrate safe-targeting ...

... without scrolling up and down the page.  Figure 3: The Education & Tools menu Further down What happens...straight from the new search bar, but the menu just above it can help you quickly and easily navigate to educational...down? You’ll find Blugene, as well as information about Addgene, our mission, links to our deposit, request...

...:02 : Niroshi and Ben introduce themselves 2:03 - 3:22 : How did you end up becoming a graduate student...graduate school community to help keep you excited about coming into lab even when things get tough in the...