...potential beneficiaries. Collaborative therapy For the most efficient outcomes, collaborative therapies must be... of the tolerance-producing antigens enunciated above. Finally, signals released from the microenvironment...are in their clinical trial phase. In all of the above therapeutics, techniques like genome engineering...are beginning to see the fruits of many years of labor with up and coming immunotherapy treatments. As ...clinic, physicians and researchers are learning much about how treatment regimens can be improved and how the...ahead of the disease in patients and developing collaborative treatments will go a long way in combating this...Additional Resources on the Addgene Blog Learn About Our Cancer Collection Pages Plasmid Grant to Aid...

...spectrum is located in a range of wavelengths of about 380–700 nm. Photons of lower wavelengths are higher... 650 nm, APC-Cy7's global excitation maximum is about 750 nm. If we were to use a laser that excites at...optimal excitation happens at higher wavelengths of about 750 nm. Source: https://www.bdbiosciences.com/en-us... we start building your first panel, we'll talk about one last principle of color emission. As you can...our blog post on color compensation to learn more about them. Alright, let's build your first panel! Designing...Conclusion You made it! In this post, you learned about the principles behind fluorophores and their colors...this article again. In the beginning, we talked about the relationship between a photon's wavelength and...

... electrophysiology data too but that’s what collaborators are for). For example, if you study neural pathways...intended brain region and/or cell type (I’ll talk more about this in detail later). But what are the different...of your opsin, but keep you eyes open for posts about the next steps in the coming weeks. Expression of...Addgene. See the Deisseroth lab homepage for details about types of constructs available, instructions for ...1, 2, 4]. And, of course, the best way to learn about the many different opsin variants and how they can...point.  In brief, some standard opsins (see table above) include Channelrhodopsin2 (Chr2), a blue-light ...construct. Stereotaxic surgery  As mentioned above, AAV opsin constructs are injected directly into...

... elevator pitch slightly if you know a bit more about the other person. If you have a poster or talk at...seemed confused, bored, or particularly excited about? If you tried tweaking your pitch throughout the...conference If there’s a cool new method that you learned about at the talk, bring it to your next lab meeting. ...questions. If you or your lab identifies a potential collaborator from the meeting, reach out to them, pointing...their talk at the meeting and why you think a collaboration would be beneficial. The poster session Before...conference. Networking All of the things we've mentioned above could be considered networking, but here are some...you remember who they are and what you’ve talked about. After the conference Follow up with those that ...

...larval stage, Drosophila will pupate, which takes about one week from the initial deposition of the fertilized...there are many resources to help you learn more about fly biology and fly-based experimental systems. ...resources will direct you to the latest information about gene function, while others offer fly stocks and...and other reagents. If you’re ever confused about the best way to study YFG in flies, there is likely an...absolutely essential. You can look up information about your fly gene using the search box in the upper ...In the next post, you’ll learn the fundamentals about how many of these resources work. Many thanks to...Site Specific Recombination in Drosophila Learn about Cre-Lox Resources on Addgene.org Find Drosophila...

...Foundation's Linda Kahl explains the Addgene collaboration this way: "Addgene is one of the leaders in ... our core values." Addgene spoke with Endy more about the BIOFAB and BIL gates plasmid kits, the Biobricks...reference to George Boole who in 1854 was thinking about logic and analyzing human language and observed ... freely available? Endy: When we start thinking about languages, humans have two types of languages: one...does; it's free to use and that tends to be true about most successful human-to-human languages. Who owns...signals. Let's say I wanted to detect every primary metabolite in a cell. The only way to do that at scale right... what can you use biology to make? If you think about the BIOFAB collection and really dig into its engineering...

...share. I spend a lot of time telling scientists about Addgene’s repository, and explaining how to deposit..., lab managers, and grad students. We’ve talked about their hot-off-the-presses papers, the awesome new... learned along the way that a connected and collaborative community makes not just for a better day at...technologies and breakthrough research. Insights about how to navigate through all stages of your career...cloning and plasmid-based technologies. Commentary about scientific sharing and the open-access movement....So join the conversation! Do you have a question about culturing iPS cells? Confused by which system to...

...have been written about CRISPR alone, and Zimmer himself has written entire books about optogenetics. Defining... been thinking about a long time. For the scientists who like to learn everything about one specific topic...wisely makes no such attempt. He is transparent about his own biases--many of which will be shared by ... a massive, expensive project that required collaboration from scientists around the world. The paper ...As a scientist who enjoys learning a little bit about many different things (part of the reason Addgene...

...SLIC cloning Learn about Gibson Assembly Additional Resources on Addgene.org Read about Choosing a Molecular...this type of primer does not provide information about the orientation of the insert. Orientation-specific...Orientation-specific primers: If you need information about insert orientation, then you might consider designing orientation-specific...insert Backbone-Specific Provides information about the size of the insert and if it's in the plasmid...plasmid dependent Doesn't provide information about the orientation of the insert Orientation-Specific... before pipetting them onto the gel. The figure above summarizes generalized expected results for the ...

...common cancers like lung (about 224,000 diagnosed this year) or breast cancer (about 246,000 diagnosed this...Mesothelioma Cancer Alliance. We have learned much about the causes of cancer and the different avenues that... asbestos exposure to the time of diagnosis) is about 35 years, but it usually ranges from 20-50 years... an orphan cancer; it is rare disease with only about 3,000 new mesothelioma patients diagnosed every ...from mesothelioma several years ago – recently collaborated with Senator Barbara Boxer to introduce the ...Additional Resources on the Addgene Blog Find out More About Cancer and Addgene's Cancer Collection Pages Learn...

...noticed Holly Bik live-tweeting about topics I needed to know more about. I caught up with her outside ...me for coffee and spent an hour with me talking about the tools I could use, and some experimental hurdles...other folks at the meeting. I didn’t know anything about these people at the time, but I met some very accomplished... my mind because we’d had discussions in my lab about whether or not to keep DNA sequencing singletons...singletons out, and there is healthy debate online about this (our lab keeps singletons). I felt like anyone...scientific conferences Read our ECR Toolbox blog post about using social media as a scientist ...

... the world. As an organization that directly collaborates with NIH-funded researchers and utilizes grants...our own initiatives, Addgene is deeply concerned about these developments. These abrupt changes threaten...valuable when budgets are constrained. This collaborative approach eliminates redundant work, saving time...partnerships: We're seeking and expanding our collaborations with industry and public and private partners...challenges. By strengthening our commitment to collaboration and resource sharing, we can navigate this difficult...collective response. Addgene stands ready to collaborate with partners across the research ecosystem to...

...Institute on Aging in a commentary about the new work published in Cell Metabolism. But, they note, there is plenty...diseases.” I don’t know about you, but I think I could use a good run right about now. And, if you are looking...be important to identify the key proteins and metabolites that are dynamically secreted by muscle during... Skeletal muscle PGC-1α1 modulates kynurenine metabolism and mediates resilience to stress-induced depression...

...but be sure to let us know if you'd like to write about your plasmids in a future blog post. No time to ...pooled libraries, but thanks to a deposit from the laboratory of Dr. Ji-Long Liu at Oxford University, we are...you plan to use GMAP for your experiments, think about depositing your new plasmids with Addgene to help...and context-dependent human fitness genes, the laboratory of Jason Moffat at the University of Toronto ...effectiveness of their library design, Moffat laboratory members conducted multiple screens in different...This work has been published in Cell. The Moffat laboratory maintains a website for the TKO library that ...selection markers, allowing for use with both laboratory and prototrophic strains. The vectors were based...

...her better! How did you first hear about Addgene?  I heard about Addgene from Joanna Brownstein, an Addgene...Additional resources on the Addgene blog Learn more about about Addgene in our "Inside Addgene" blog series Subscribe...money for childhood cancer through the sale of art. About two years ago, I had a big role in organizing an...making, reading, sailing, and gardening. Learn more about Julie Cicalese  Ms. Cicalese served as the Chief...

...variety of short- and long-form written content about timely scientific topics, as well as archived seminar...members gave up a beautiful summer Saturday to learn about science, but they didn’t seem to mind! Attendees...consistently mentioned how pleased they were to learn about these exciting scientific topics at a level accessible...other attendees and getting the chance to hear more about what life is like as a student researcher. The future... with the community? Addgene would love to hear about your efforts to engage the public - let us know ...know in the comments section! More information about SITN: DayCon event website (includes archived content...

... Learn about research applications of antibodies in Introduction to Antibodies Learn more about fluorescence...Stay tuned for a future blog post to learn more about these techniques!  Direct vs. indirect immunofluorescence...different approaches to be familiar with when talking about IF, direct and indirect. For direct IF, researchers...immunofluorescence procedure with key considerations about each step. Image created with BioRender.com ...of protocol you will need to use. (To learn more about antibody validation, this commentary by Uhlen, et...Fluorescence Microscopy Techniques is Best for Me? Read about another common antibody application in The Basics...

... 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... Video production, just like science, is always about iteration and improvement. You have an idea, you...

...Something that I struggle with as I’ve been thinking about my career options is being an overthinker. Being...Stepping into this new virtual world, I was worried about how my ideas would be translated through Zoom calls...Series on Instagram. I interviewed fellow Addgenies about their role in the company. One of the things I wanted...Specialist. Both of these Addgenies really made me think about my future. They made me rethink having a structured...not only in terms of its employees but in general about anyone who steps foot through their door.  In my...just an intern but I was an Addgenie. I thought about all that could go wrong with this internship but...

...shown in photo), has been speaking out and writing about issues in our science training infrastructure for...some years.  He spoke eloquently and vehemently about the need for change.  Dr. Bourne strongly believes...fact that the PhD and Postdoc advisors only know about academia and alternate training needs to be provided...have to be in academia to make change.  He talked about his own unique way of funding basic research at ...at NEB and got some applause when he talked about how he boycotts publishing in all but open access journals...postdoctoral program.  His strongest comments were about the astounding lack of reproducibility being recognized...

...be used in people until we learn more about safety and more about the how to use genome edited stem cells...because Jennifer did her graduate work one floor above mine in the lab of Dr. Jack Szostak, who went on...ceremony honored Charpentier and Doudna, who talked about their research leading up to the discovery of the...Charpentier met at a 2011 conference and decided to collaborate on what was a “curiosity” driven project. Both...importance of the strong, cross-disciplinary collaboration they shared to the work they did together. They...

... outreach and community engagement. Information about contacting Addgene can be found on our Contact page...Europe office located in London and operated in collaboration with LGC Standards. And now you’ll be able to...ll be poised and ready to answer your questions about Addgene, plasmids, and depositing (all while our...carrying out my studies, and getting them from collaborators was extremely helpful, but with hindsight I ... and Australia, viewing science as a global collaboration. This is why she is excited to be working at...This love of interacting with people and talking about science makes her the perfect Addgenie and she can...

...Additional Resources on the Addgene Blog Learn about Our Collaboration with Fellow Nonprofit Seeding Labs Addgene...the American Cancer Society, or maybe you think about a local food pantry or community arts organization...community Addgene fosters a culture of respect, collaboration, and fun Addgene believes there is strength ...accelerate research with easier sharing and collaboration. Joanne and all the Addgenies: Adam, Amanda ...Connie Cepko Resources on Addgene.org Learn More About Our Mission Meet the Addgene Leadership Team Get...

... post if you debating about an academic or non-academic career Learn more about alternative science careers... written for selfish reasons: I wanted to learn about careers in science communication. When I started...Communication Internship with Addgene, I didn’t know a lot about scicomm, but had enjoyed writing a few Addgene guest...option. So if you’re interested in learning more about science communication careers, you’re in the right...careers here Resources on Addgene.org Learn more about careers at Addgene ...

...Reddit AMA  Few of us spend much time thinking about conserving resources in the labs. Disposable plastic...Dudnik, Founder of Seeding Labs, tells the story about working as a student in a rice lab in the Ivory ...didn’t feel the need to reuse plasticware or think about conserving reagents and buffers.  However, I had...environmentally conscious friends who had me thinking about energy conservation even before it was a popular...Turns out, one could make a lifetime supply for about $10 US dollars worth of glass powder pruchased from...So, I designed my own gel box (see illustration above) and bribed the guys in the hospital machine shop...

... Delivery Methods Learn about Mouse Work on Exercise and Depression Read About Brainbow Resources on ...success is not always guaranteed. To learn more about this process check out our Mouse Modeling blog posts...by CRISPR. The sequence started clean and then, about 100 bps into the run, it got “dirty”, with multiple...makes use of 3 components, a guide RNA (gRNA) of about 125 nt that specifies the target, the Cas9 endonuclease...by Southern blot. From the efficiency discussed above extends 4 major advantages of CRISPR mouse editing...targeting. It also costs less. With CRISPR, it may cost about $5,000 to generate founder mice, while with conventional...

...mentoring group (see above). However, now that we are all so good with video calls, how about trying your hand...blog or podcast, initiated and maintained a collaborative project, served on the board of a professional...appointments with people all over the world. Think about contacting alumni of your lab or university or use...suggest you speak with. See this post that’s all about informational interviews, for more details on how...progress on both building relationships and thinking about future career options. 5. Write a grant - even if...

...the Addgene blog Learn more about basic viral vector elements Read about four considerations for the ...Key elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal...routine plasmid propagation, with complete ITR loss abolishing AAV packaging. To prevent ITR loss, it’s best... Other cis-regulatory elements Next let’s talk about some of the elements that aren’t required parts ...the perfect AAV tool Learn about synthetic promoter AAVs for cell-type expression in retinal cells Resources...

...This great resource allows the undecided to learn about a variety of science professions based on their ...training focuses on using multidisciplinary, collaborative, and translational approaches for scientific...skills. Although the word “extracurricular” may be taboo in the lab, multiple research studies highlight ...social media (Twitter, Facebook, etc.) to learn about available jobs, fields, and trends “Never Leave...from your lab bench.  Want further information about shaping your career to your values, interests, and...Let us know what topic you'd like to learn more about next. Leave your suggestions in the comments section...

...student at MIT. While I had always enjoyed learning about biology in high school, it wasn’t until I started...college that I really fell in love with it. Something about being able to actually hold all those seemingly ...seemingly abstract biological reactions that I learned about from textbooks just made everything click in my head...students build circuits in physics class to learn about electronics or mix chemicals together in chemistry...a power source, but now you don’t have to worry about operating the rest of the car as well. Similarly...with us! We’ll work with you to get the word out about your plasmids and their potential educational uses...

...”) of DNA, have changed our collective thinking about the feasible size and scope of projects in many ...next generation (NGS) sequencing have prevented laboratories from using these methods for routine processes...daunting for large numbers of samples. As mentioned above, one of the primary technical benefits of plexWell...1ng – 100ng) are normalized to two-fold variation above and below mean read count per sample, and the amount...NGS readout also reveals additional information about the presence of non-plasmid DNA or heterogeneous...   Check out the SeqWell website to learn more about our plexWell kits and services! Many thanks to Joe...

...Learn About our Cancer Colleciton Pages Use the ClonTracer Library for Lineage Tracing Learn About Cancer... For many years, we’ve hoped that large scale laboratory-based functional experiments that classify which...Collection). In addition, we worked with many collaborators to aggregate a sequence-confirmed series of ..., we used the cancer plasmid collections listed above to demonstrate how gene expression profiling, systematic...with the data generated from these alleles in the above publications), it will be possible for many others...

..., we — Jo, Brook, and Angela — most often think about accessibility in terms of the content we write for...provide visually impaired visitors with information about images that a sighted reader would glean from the...information in the paper.  Of course, when you think about writing an entire scientific paper, you may feel...done? What are we trying to fundamentally change about their knowledge?    Here, this image is being used...extract virus from the correct layer.  Third, think about the venue you are using to present the image and...participants to do the same thing, and even though only about 10% of them had heard of alt text before, they all...

...more about their needs and any problems that they are trying to solve. After learning more about the stakeholders...the company and then go to grad school. However, about a year and a half into my role, I realized that ... my product management (PM) career. Had I known about product management earlier on in my career, I’d ...interests. As such, I want to be able to share more about this nontraditional career path for others who love...management.  A day in the life  Now that you know a bit about what a software product manager does, you might ...

...for Scientists Series. I have been thinking a lot about Mentoring for over 10 years. Many successful scientists...Medical School researcher Connie Cepko to learn about her mentoring style. While working on this project...postdocs I have started to talk to trainees more about peer mentoring. They want to find mentors to help... questions I get is, “What are we going to talk about?” There are an infinite number of ideas on the internet...from a lab, career or industry of interest – learn about their career paths and get their advice. Attend ... your own mentoring group or with any questions about mentoring. Read the other posts in the series to...

...work if both parties are diligent about replying and thinking about good questions and answers. Professional...Sometimes, it is not who you know but who knows about you. I found some other great tips for finding mentors...Medical School researcher Connie Cepko to learn about her mentoring style. Mentors at home and away We...Catalyst Report “Making Mentoring Work” to learn about what makes a good mentoring program. If your organization...much less threatening! If they say no, don’t worry about it. It is a compliment to ask someone to share their...

...biosafety cabinet with cloning rings or following labor-intensive limiting dilution protocols? If this sounds...fluorescent microscopy must be used. To learn more about the most suitable fluorescent microscopy technique...a laser and the instrument collects information about the cell’s size, complexity and fluorescence intensity...captured by detectors and provides useful information about the cell. Light that continues in the same direction... forward scatter (FSC) and provides information about the relative size of the cell; larger cells produce...called side scatter (SSC) and provides information about the cell’s complexity. A cell with a high degree...other methods using fluorescent proteins Learn more about antibodies Resources on Addgene.org Visit the ...

...is referred to as an antigen. You can learn more about antibodies in our Antibodies 101 animation, What... expressing this protein. Read on to learn more about antibodies and how to use them in the lab! Parts...lots is a big concern. You can find more details about polyclonal antibodies on our Antibodies 101: Polyclonal...not necessarily what you are getting. Learn more about monoclonal antibodies here. Recombinant antibodies... as well as compare between samples. Learn more about flow cytometry. Cell and tissue labeling Antibodies...Additional resources on the Addgene blog: Read about another reagent sharing platform for antibodies,...Developmental Studies Hybridoma Bank Learn more about nanobodies Watch our Antibodies 101 animation, What...

... careers, which she hadn’t.  He then said, “How about being a science writer?” to which Susan replied,...replied, “What’s a science writer?” Learning about SciComm by doing SciComm Writing had always been a strength...the National Eye Institute (NEI), where she wrote about NEI research for a general audience. She then got...always plenty of writing: plain-language stories about Fred Hutch research, pieces for donors such as fundraising...articles are just harder to write. “There’s something about being a writer that can be a very painful process...using social media as a scientist here Learn more about storytelling in science ...

... the best way I’ve come up is this: It takes me about the same amount of time to read a 100-page genre...it instead as valuable feedback.  Listen. Think about the questions you’ve received, read the comments...aware of cognitive load changed the way I thought about engaging with learners. If something I was communicating... instead of blaming the audience, I would think about how I was presenting the information. Maybe I needed...observe and adjust on the fly.  Once you can think about everything you’re asking your learners to do, it... engagement with your target audience. Thinking about cognitive load will help you understand why you’...

... time, when someone speaks about an affinity reagent, they’re talking about a standard antibody: An ~150kD...mutation, allowing diabodies to be used for assembling protein nanostructures. Diabodies readily co-crystalize...for clinical applications, including imaging.  Diabodies are ~60 kDa proteins formed by two Fab fragments...2019). Structural diversity and flexibility of diabodies. Methods. 10.1016/j.ymeth.2018.09.005 Muyldermans...

...silence, I opened my mouth to panic-babble something about Western blots that made no sense, even in my head...doubt anyone but me remembers any of the incidents above–and the dumpster noises were just last week!  Nerves...attention? fear of messing up?–and use the three tips above to make a strategy that can help. Even something...speaking as a transferable skill The worst thing about doing public speaking as a scientist is that you...perfectly, you truly are a rock star.  The best thing about public speaking as a scientist is that you will ...increases visibility of your work, opens up collaboration and growth opportunities, and gives your entire...

...20th anniversary was about all the amazing years behind us, as much as it as about the amazing years in...Lunenfeld-Tanenbaum Research Institute, posting about Addgene’s 20th anniversary on Twitter. In the beginning...Julien Béthune, Professor at HAW Hamburg, posting about his Blue Flame Award on Bluesky. In 2016, we also... As the pandemic slowed, Addgene began to think about how we could expand our support for the research...part of the Neuroscience AntiBody Open Resource (NABOR) platform, supported by an NIH BRAIN Initiative....

...differently based on their audience. How you talk about your experiments to your PI, your colleagues, or...have cross-team projects within your own lab, collaborations with other scientists within and outside your...not happen in the lab, they have parallels to collaborative lab work. Corralling context and junking jargon...difficult to know what kind of plate they were talking about during the meetings. And “plate” certainly wasn’... a lot on context. New people in the lab or collaborators will not have the context you do, which means... Additional resources on the Addgene blog Read about other transferable skills Find all of our science...

...Individual Development Plan Learn a Little about The Academic Labor Environment from the Future of Research...research.  It helps to spend some time thinking about what your goals are for the coaching relationship...ask for references and follow up with 3-4 to hear about those clients’ experiences. Good coaches will have...with someone you connect with. Talk to the coach about their “style” and make sure this is a style that...coaching group is Still Point Coaching. One nice thing about a group is that sometimes this offers the option...

... work into finding robots that’ll take a little labor off our hands. Good news! There is probably a machine...96 transformations in one run compared to doing about 50 transformations in the same amount of time by...machinery we have been able to transform 96 samples in about 4 hours with a greater than 95% success rate! Automating...plates in just under 3 hours or one 96 well plate in about an hour. Automating vial fiIling The majority of...35 minutes compared to a lab assisstant filling about 200 vials in 45 minutes by hand. How can you start...on each tube by hand. We’ll be sure to tell you about more lab updates as we develop them. In the meantime...

...ask the depositing laboratory to review the discrepancies. If the depositing laboratory confirms that these...finish the library preparation process only takes about one day, even for six plates (576 individual plasmids... we perform a 2x251 run on our MiSeq that takes about two days to complete. After the run completes, we..." In some cases, there will be more information about our sequencing results in the Depositor Comments...to note the appropriate barcode for each sample. About 24 hours later we unload the FASTQ data from our...

...Sainsbury Laboratory. She is passionate about science communications and loves talking about science, ...Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory. Are you a science student or early career researcher...break into science communication? Everybody goes about this in their own way. The career paths into science...Society gives training in media skills, writing about your research and residential communication. If ...

...distance so ideal implant placement should be about 0.4-0.5mm above the site of your stimulation. The scatter...scatters a great deal once it enters tissue, placement above the site of stimulation guarantees that the majority...instead). Laser stimulation         In the video above, Channelrhodopsin 2 (Chr2) has been expressed in...These lasers are relatively cheap and will run you about $2-3,000/laser. However, an important caveat is ...output of the laser must be controlled. As discussed above, each opsin variant has slightly different activation...

... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential...with successful recombination reported in all laboratory E. coli strains tested so far. The use of this...low cost and can be immediately adopted by any laboratory. All procedures are performed using the same... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Jake Watson is a postdoctoral...postdoctoral researcher in neurophysiology at the MRC Laboratory of Molecular Biology. He is currently investigating...García-Nafría is an investigator scientist at the MRC Laboratory of Molecular Biology. He is interested in neuronal...