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...Resources on Addgene.org Addgene’s AAV Guide Fluorescent Proteins: Biosensors Addgene’s Viral Services...calcium sensors. Selecting a calcium sensor Genetically encoded calcium indicators (GECIs) are fluorescent...neuronal cell body or the axon (GCaMP6 only). The genetic elements required for Drosophila studies have been...chemical biology. 2021 May;17(5):509-18. Inoue M. Genetically encoded calcium indicators to probe complex brain...Aug 1;169:2-8. Suzuki J, Kanemaru K, Iino M. Genetically encoded fluorescent indicators for organellar...SA, Looger LL. Imaging neuronal activity with genetically encoded calcium indicators. Cold Spring Harbor... Tsien RY. Measuring calcium signaling using genetically targetable fluorescent indicators. Nature protocols...

...share your lab protocols online Addgene Website: http://www.addgene.org/ If you’re working with plasmids...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...protocols. Since we began sharing this protocol on the Addgene website in December 2006, the pLKO.1-TRC cloning...the UK, South Korea, and beyond!   Check Out How Addgene Thinks About Developing Lab Management Software...plasmids and you decide to share them using Addgene’s repository, you can also share your associated protocols...submit an updated protocol? Just email us at help@addgene.org and we will be happy to upload your newest resources...gRNA_Cloning Vector (deposited by George Church’s lab). Addgene has also cataloged a variety of useful molecular...

...FOP until a cure is available. Addgene and AJ's Avengers This year Addgene, where my wife Lianna has made...their plasmids with Addgene and join us in the fight against FOP. Second, Addgene will be hosting the ...years. Additional Resources on the Addgene Blog Learn about Addgene's Previous Work with Rare Disease Researchers...potential FOP cure But, there is HOPE. FOP is a genetic disease typically caused by a single R206H point...receptor ACVR1. ACVR1 triggers appropriate bone regeneration in healthy patients. However, the R206H mutation...helping us fight against FOP in two ways. First, Addgene is distributing ACVR1 Plasmids from a variety of...Potentiall HIV Treatments with CRIPSR Resources on Addgene.org Find ACVR1 Plasmids Find Plasmids for Disease...

... KillerRed, a genetically encoded red fluorescent photosensitizer protein that generates ROS when illuminated...16369538. Additional Resources on the Addgene Blog Read Other Optogenetics Blog Posts Sign Up to Receive the...residues. Compared to MG, MG-2I has increased ROS generation when bound by the dL5** FAP.  The iodine-modifications...the main advantages of MG-2I-dL5** over other genetically encoded photosensitizers is that it works with...cells.  He et al also show that MG-2I-dL5** only generates small amounts of non-specific ROS when exposed...Watkins, S. C., . . . Bruchez, M. P. (2016). A genetically targetable near-infrared photosensitizer. Nature...Chepurnykh, T. V., . . . Lukyanov, K. A. (2005). A genetically encoded photosensitizer. Nature Biotechnology...

...requested from Addgene. Figure 2: Total COVID-19 plasmids requested from Addgene from March 23, ...resources on the Addgene blog Read more about COVID-19 databases and resources Learn about Addgene's work to ...all scientists Resources on Addgene.org Find COVID-19 plasmids at Addgene Browse the entire plasmid collection...2020). The authors requested these plasmids from Addgene. This publication then resulted in the development...3X-Flag-SARS-CoV-2 S, which is now easily available through Addgene and has already been requested dozens of times ...develop available to the scientific community. Addgene was awarded a Fast Grant to ensure that the scientific..., 2020 to July 27, 2020. Browse Addgene's COVID-19 plasmids and resources! Case study 3: Mouse strains...

...distances. For instance, how much force can be generated by conformational changes in proteins? How can...can be addressed using R bodies: simple, force-generating protein machines that can extend and retract ...demonstrated that R bodies can be “tuned” with mutagenesis to extend at different pH values. Using a screen...in a particular region of the R-body. Further mutatgenesis of this region produced R-bodies that could ... wild type and mutant R bodies can be found at Addgene here. What can R bodies do for you? R bodies are.... & Pond, F. R. Organization and expression of genetic determinants for synthesis and assembly of type...

...Read Other Inside Addgene Posts Resources on the Addgene Website Meet the Addgene Management Team Join...Additional Resources on the Addgene Blog Get Management Advice from Addgene Executive Director Joanne ...meetings. Make sure to keep them manageable. At Addgene, teams can add as many points as they like to the...the meeting, take notes on the agenda points. At Addgene, we like to do this live on the screen using a ...review or project plans, before the meeting. At Addgene, we like to send around a Google doc where people...provided some tips above for what works well at Addgene, but meetings are never one-size-fits all. Feel...Join the Addgene Community ...

...commercialization. Many of the genetic pieces developed over the years were generated by large agricultural companies...you have developed a genetic tool for plant engineering, please share it with Addgene to both build the ...transformation to testing and a lack of well-characterized genetic tools make it challenging to engineer a specific... biotechnologists face is public perception of genetic modification (GM) and synthetic biology. The mistrust...research – this comprehensive understanding of the genetic pieces underpinning these mechanisms – our project...limited distribution. We hope that will change with Addgene’s expansion into plant synthetic biology. There ...and we are excited to see how this exchange of genetic tools facilitates plant science. Challenge #3: ...

...Plasmids 101 eBook Resources on Addgene.org Join the community of Addgene depositors here Check out our...repeating before you can finally dive deep into generating the data you’re excited about. When we were ...intervening five years, inflation has occurred. Here at Addgene, the consumables cost per stab has increased 50%...plasmids. Use repositories Repositories such as Addgene help share materials quickly and cheaply. We save...last minute experiments or changes. Fun fact: Addgene’s high-throughput approach means we virtually always...community engagement with and use of repositories like Addgene provides large-scale savings. How large? Using ...average of 5 hours of hands-on time per plasmid, Addgene, since its founding in 2004, has saved the scientific...

...Fragments generation After you’ve designed your cloning scheme, you’re now ready to generate fragments...Paolo Colombi, a product development scientist at Addgene. Cloning can be quite an arduous process. The PCR...fragments. You can generate the parts of DNA that you want to assemble together in a combination of different...conditions. Purifying DNA fragments Once you’ve generated your DNA fragments, it is always a good habit ...step has to be strategy specific, there are few general parameters that you can adjust: temperature and...and time of incubation, and amount of DNA. As a general rule, try to use an excess of insert compared to...currently the product development scientist at Addgene. In the lab he develops new assays and processes...

...Resources at Addgene Find the ClonTracer Library at Addgene Find Lentiviral Plasmids at Addgene Check out...abundance and clonal dynamics over time using next generation sequencing (NGS). The library was developed when... al., 2016).  Bhang deposited the library with Addgene because she wanted to make it as easy as possible...and she is excited to deposit the library with Addgene because it is a “really great way of broadly sharing...sharing these kinds of reagents.” Here at Addgene, we’re excited to see the creative new ways you put ...and for providing this powerful resource to the Addgene community! Carrie Bhang is a research investigator...in applying what we have learned about tumor heterogeneity to drug discovery and the development of combination...

...fit in a freezer). Our Addgene celebration was so much fun! Here's the Addgene Leadership team in all ... a million plasmids would fill? Two weeks ago, Addgene edged passed our one million plasmids shared milestone...might have you wondering how many plasmids leave Addgene headquarters every day. That would be an average...our shipping room above), but as Joanne Kamens, Addgene’s executive director, said during our celebratory...it takes to make one plasmid and how much time Addgene has saved researchers. Assuming 5 hours of hands-on...Addgenies on social media or around the office. @Addgene congrats for 1 million orders. @RothLabUNC accounts... — BryanRoth (@zenbrainest) August 21, 2018 As Addgene grows, we have been able to adapt to the needs ...

...proteins for optogenetic control of CRISPR-Cas9. By combining AcrIIA4, a Streptococcus pyogenes Cas9 inhibitor... a few of the new CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR...adapted for detecting SNPs. Learn more from this Addgene blog post Read the paper in Science Find the...the Cas14 plasmids Anti-CRISPR proteins for optogenetic control of CRISPR-Cas9 Anti-CRISPR proteins that...editing is now possible. Learn more from this Addgene blog post Read the paper in Nature Methods Find...a new CRISPR tool you’ve recently deposited to Addgene and you’d like it to be included in the next What...please let us know! Additional resources on the Addgene blog Download the CRISPR 101 eBook Find our CRISPR...

... plasmids for generating your own libraries are also available in the Sybody Generation Toolbox plasmid...lab found a way around this by using plasmids to generate sybodies, synthetic nanobodies against the spike...blog post.  A rapid selection platform for generating sybodies against membrane proteins Developing...membrane protein of interest using the Sybody Generation Toolbox and commercially available reagents in...libraries (Zimmermann et al., 2018). Find the Sybody Generation Toolbox! Starting with libraries and a biotinylated...relevant plasmids are available in the Sybody Generation Toolbox kit. Find the Seeger lab's SARS-CoV-2... S, Dawson RJP, Geertsma ER, Seeger MA (2020) Generation of synthetic nanobodies against delicate proteins...

... Figure 1: Discussing next-generation sequencing at the Addgene bioinformatics journal club! ...and level of general busy-ness will most likely the primary drivers of frequency. At Addgene, we opted to...recurring journal clubs on Immunology and Microbial Pathogenesis that occurred once a week for several months...want to pick a topic and have no one show up. At Addgene, we wanted to think more closely about the wealth...wealth of high-throughput sequencing (HTS) data we generate and maintain. It became clear that while several...messaging but we as scientists often prefer data. At Addgene we rely on polls and surveys. A mix of closed (...wandering monologue?  Assuming the feedback is generally positive and the meeting seemed to be of use, ...

...training in a variety of transferable skills. At Addgene we're helping to fill this gap by both helping ...yourself in a creative rut, read on!   Fig 1: Blugene using creative problem-solving in the lab   ...to do this and are still struggling with idea generation, try this thought exercise: Consider the issue...three-ring binder as a starting point, the ideas generated were hardly novel. When told to use a roller skate...that way— but the idea behind the study holds. To generate novel ideas, start with an out-of-the-box concept...option for any meeting in which the goal is to generate unique ideas. Recognize ideas can come any time... had over lunch are just as valuable! In 2019, Addgene implemented a company-wide New Ideas Survey. Anyone...

...Resources at Addgene.org Find careers at Addgene Watch out career videos Sign up for the Addgene quarterly...making this transition. In this installment of the Addgene career guide, we’ll think about what causes this...job candidate for a senior scientist position at Addgene, the skeptical applicant asked if she would partake...of culture shock above, but after polling some Addgene colleagues, we’ve come up with a list of things...the years that one of the things I enjoy about Addgene is that I’ve never been bored. There is always ...someone who coordinates cross-team projects at Addgene, I find that my vocabulary improves on a weekly...fix them, and take a look at these tips from the Addgene blog. You’ll have the opportunity to discover...

...nature18308   Additional resources on Addgene.org Addgene’s General Use Antibodies Antibody guide Antibody...applications. You can find FLAG antibodies on Addgene’s catalog including a mouse monoclonal (M2) as well... acid peptide (EQKLISEEDL) derived from the protooncogene of the same name. The tag has a net negative...clone used today! Goat, chicken, mouse, or rabbit? Addgene has you covered with monoclonal antibodies to Myc...both cleave after the DVPD sequence. Check out Addgene’s monoclonal antibodies (mouse, rabbit, and chicken... antibodies or guidance on designing a GFP tag Addgene has you covered!   Potential tag issues Tags have...Antibody protocols Additional resources on the Addgene blog Antibodies 101: Epitope availability Plasmids...

...Resources on addgene.org  Addgene's Viral Vectors Guide Addgene's Chemogenetics Guide Addgene's Optogenetics... our collection, on the Addgene eBook webpage.  More resources on the Addgene blog CRISPR 101 eBook en...

...Blotting Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene ...an IF experiment. What is immunofluorescence? Generally speaking, IF is an application in which antibodies...iBiology). IHC and ICC are essentially just more general terms that can also refer to assays using non-fluorescent... visualize their target. These antibodies are generated against a specific antigen and then conjugated... more common is that secondary antibodies are generally polyclonal, which means that multiple secondary...amplification. Another reason is cost effectiveness. Generating and validating primary antibodies is a time consuming...antibodies that exist and you can begin to see why generating fluorophore-conjugated versions for each primary...