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...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...

...zebrafish, and mice. Second Generation iGluSnFR: SF-iGluSnFR In the second generation of iGluSnFR, the Looger...being deposited at Addgene. Other plasmids and pAAVs can be ordered directly from Addgene. The Podgorski ... for Synaptic Imaging However, the first two generations of iGluSnFR had limitations: they were not optimal...extrasynaptic glutamate release. Building on the previous generation of the SF-Venus-iGluSnFR (itself an improvement...IPTG induction to express soluble protein CAG: Generic strong promoter. Formulated as AAV-DJ for neuronal...References and Resources Additional Resources on the Addgene Blog Learn about other Fluorescent Biosensors ...Fluorescent Protein Featured Topic Page Resources at Addgene.org Find Fluorescent Protein Biosensors Browse All...

...of sample preparation for high-throughput next generation (NGS) sequencing have prevented laboratories ...As part of our mission, we’ve been working with Addgene to develop and apply our plexWell™ Library Preparation... for NGS-based sequencing and confirmation of Addgene’s large and growing collection of curated plasmids...are using this technique in our partnership with Addgene to sequence large numbers of plasmids. plexWell...does not vary by DNA input amount. plexWell™ + Addgene = high quality NGS-assembled plasmids Because of...information about the presence of non-plasmid DNA or heterogeneous and/or mixed clones present in the original ...original sample. At seqWell, we have worked with Addgene to develop a highly optimized bioinformatics pipeline...

...Before I started writing for the Addgene blog, sharing Chemistry Cat memes was how I used social media...social media as a scientist. Now I write for the @Addgene blog & I'm my lab's resident #scicomm expert. What...using social media as a scientist started with the Addgene blog. I wrote my first blog post out of boredom...person in the lab. When I saw a request for an Addgene guest blogger post about colony PCR, I signed up...and accepted a position to work remotely as an Addgene Science Communication Intern. Developing science... using Twitter, I found this paper. I wrote an Addgene blog post about it and shared the paper with my... this panel, but my consistent writing for the Addgene blog (~1-2 blog posts per month) and use of Twitter...

...Additional Resources on the Addgene Blog Read Additional Cancer Blog Posts Addgene's Cancer Collection Pages...this multi-pronged attack. Inflammation and tumorigenesis The conference opened with a very interesting... between tumor and immune cells in real time. Addgene depositor Professor Max Krummel from UCSF showed...engineering (Ref 5)  as described by Blue Flame Addgene depositor Professor Michel Sadelain from the Memorial...poster is pTomo-Ras which has been deposited at Addgene was used to produce the glioblastoma model for ...addition, it was exciting to see that the use of optogenetic tools is expanding to immunological research....with the scientific community, contact deposit@addgene.org to find out more about our free deposit and distribution...

...Addgene is excited to announce we now offer an export license application for rabies materials! Some ... export-controlled materials in the USA, where Addgene is located.  The export license, if granted, allows...ship rabies plasmids and viral vectors containing genetic elements from rabies plasmids to international ... export control license, once approved, allows Addgene to ship plasmids or viral vectors containing sequences...project when purchasing export-controlled materials. Addgene will then apply for the license for distribution...processing costs. Once the application is approved, Addgene then ships the material following our normal process... If you have any questions about this or other Addgene shipping and ordering processes, don’t hesitate...

...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.      Additional... Today on the Addgene Podcast, we have another interview conducted by European Outreach Scientist Benoit...creation of improved fluorescent proteins through mutagenesis. Chapter 6 - The future of FRET and fluorescent...Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes. You can follow him...Additional Resources from the Addgene Blog Visit our Fluorescent Proteins featured topic page Read Joachim's...Subcellular Structures and Organelles Resources at Addgene.org Find Fluorescent Protein Plasmids Find Plasmids... addgene podcast...

...resources on addgene.org  Antibody GuideRecombinant antibodies available from AddgeneBrowse our recombinant...I love Addgene's new Antibody Guide and not just because I helped create it. But while antibodies, as...space to give proper credit to. Thankfully, as Addgene's resident blog editor, I could take the needed ...alternative reagents are created in the lab, instead of generated in vivo, they have higher reproducibility and ...small enough to be produced in bacteria and are generally cheaper to make than antibodies. Minibodies are...Designed ankyrin repeat proteins, or DARPins, are genetically engineered small affinity proteins, around 14...instead of the antibody constant region. They are generated using combinatorial libraries, and high specificity...

...plasmid-based antibodies Resources on Addgene.org Discover Addgene’s antibody plasmid collection ...antibodies are produced Polyclonal antibodies are a heterogeneous mixture of many antibodies that recognize the... response.   Polyclonal antibodies are generated by injecting an animal with an immunogen, then...weeks later. Created with biorender.com. To generate polyclonal antibodies, an animal (such as a rabbit...might be the full-length protein that you want to generate antibodies against, or a unique peptide sequence...experiments to be aware of signal strength. In general, polyclonal antibodies produce a higher level of...the same species your polyclonal antibody was generated in or a polyclonal isotype control, to account...

...the blog Resources on Addgene.org Find fluorescent protein plasmids at Addgene Find chromoprotein plasmids...a redo :-( @SandTBioSci #CompetitiveStreaking @Addgene pic.twitter.com/mk44J4Gcj9 — Dave W will get vaccinated...found the chromoprotein plasmids available from Addgene, he knew it would be a perfect fit for the streak...biology. “What I'd like to do with the plasmids from Addgene is have the students transform them into different...transformation experiment, he plans to show how different genetic backgrounds of the host strains affect transformation... Youtube:     Additional resources on the Addgene blog Read our other blog posts on fluorescent ...plasmids at Addgene Check out additional written and video protocols ...

...soon with some brand-new Addgene swag!   Last month, we introduced you to Blugene and Aavery's newest friend...friend, our Addgene antibody mascot, here to help us with our upcoming affinity reagent distribution launch...this form or head over to Twitter to vote on @addgene's Twitter poll. Voting is open until Feb. 22, 2022...winning name on Feb. 24. Stay tuned!    Original Blugene and Aavery would like your attention, please, as...that has led them to their biggest discovery yet: Addgene will soon be distributing affinity reagents. Name...hashtag #AntibodyMakesThree on Twitter. As with Blugene and Aavery, we’re looking for a gender neutral ... four will receive some rockin’ newly designed Addgene swag. Good luck and happy naming!   Fig....

...breaking into more audio and video on the Addgene Blog and Addgene website. As we push forward with these...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...present plasmid info with a new segment on the Addgene podcast - The Hot Plasmids Segment. Click on the...Listen to the entire Hot Plasmids episode! We generally publish 5-10 Hot Plasmid articles with each of...Additional Resources on the Addgene Blog Listen to Our Podcast Interview with Eye Researcher Connie Cepko...Science Videos from Tom McFadden Resources at Addgene.org Sign up to Receive Our Newsletter Check Out ...

... Brian O'Neill.  More resources on the Addgene blog Addgene's Data Hub What is a Virus (and a Viral Vector...Addgene’s viral vector services allow scientists to conduct experiments faster, skipping the preparation...Table 1: New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor...serotype pAAV_hSyn-PdCO-mScarlet-WPRE AAV1 Optogenetics Ofer Yizhar New viral tool pAAV-Ef1a-fDIO...pAAV-Ef1a-fDIO hChR2(H134R)-EYFP AAVrg Optogenetics Karl Deisseroth New viral tool AiP13987: pAAV-AiE0452h_3xC2...different serotypes. Not sure what to request? Addgene's Data Hub can help, with over 100 user-submitted... the link below to keep up with the latest in Addgene's viral vector services and resources!   This blog...