...structure-guided direct evolution, Nathan Shaner and colleagues from The Scintillon Institute, Allele Biotechnology...Clavularia sp. protein CFP484. Hui-wang Ai and colleagues from the University of Alberta and University... in a 2.6-fold increase in signal relative to mCerulean imaged with a standard CFP set. mTFP1 can be used...would like to share with the scientific community, please deposit the plasmids at Addgene! References Ai... to the Addgene blog Resources on Addgene.org Learn about the Michael Davidson fluorescent protein collection...

...Foundation, ABRC’s outreach program kicked off with the release of 20 education kits designed for use in K-12 and...ABRC database and ordering platform soon to be released. For individuals interested in ordering from ABRC...has many benefits. Engaging students in active learning through hands-on experiments allows them to develop... out opportunities to engage people in science learning. Whether visiting classrooms, hosting field trips....     Additional Resources on the Addgene Blog Learn how BioBuilder can bring Synthetic Biology into ...

...Resources at Addgene.org Learn about Popular Plasmids in the Repository Learn about New Plasmid Technologies... relatively low efficiency of biotinylation, at least in vitro. Our solution was to exploit the best of...proteins that lacked TEV. Although we were unable to cleave TEV within the protein we tested, it may be possible...plasmid in NEB T7 SHuffle bacteria (C3029J). We are clearly NEB “fanboys” as the bacteria significantly enhance...

...contributed by guest blogger Fredrik Wermeling, leader of a research group at the Centrum for Molecular...design gRNAs for all the purposes discussed above. Please see videos and text on the website for more detailed...have been “green listed” by several algorithms. Please read the “Detailed Information” text linked to ...and libraries using strategies related to other nucleases as well as inhibitory and activating approaches...PMC5292764. Additional Resources on the Addgene Blog Learn about CRISPR Screens How to Design Your gRNA Tips...for a 1st CRISPR User Resources on Addgene.org Learn More About CRISPR Find CRISPR Screening Libraries...

...with a microbe, to trigger immune cascades that clear these infections. Anything that generates an antibody...immune system is referred to as an antigen. You can learn more about antibodies in our Antibodies 101 animation...isolate cells expressing this protein. Read on to learn more about antibodies and how to use them in the...produce is not necessarily what you are getting. Learn more about monoclonal antibodies here. Recombinant...simultaneously as well as compare between samples. Learn more about flow cytometry. Cell and tissue labeling...antibodies, the Developmental Studies Hybridoma Bank Learn more about nanobodies Watch our Antibodies 101 animation...

...excitation and emission spectra. Both spectra possess clear maxima where excitation and emission are the most... 500 nm and a global maximum at around 560 nm, leading to different levels of brightness, depending on...check out our blog post on color compensation to learn more about them. Alright, let's build your first...cell culture, the transduction process itself also leads to some cell death. As a result, there is a high...   Conclusion You made it! In this post, you learned about the principles behind fluorophores and their...this process creates visible colors. Next, you learned about how fluorophores can be classified into different...

...attempted to study cancer from every possible angle to leave no stone unturned, but cancer has been producing...between tumors and the immune system We’ll begin by learning how the immune system naturally interacts with...Through interactions with the immune system, tumors “learn” how to escape it. In this process, cancer cells...tolerance-producing antigens enunciated above. Finally, signals released from the microenvironment help create tolerance...enter the clinic, physicians and researchers are learning much about how treatment regimens can be improved...PMC3433802. Additional Resources on the Addgene Blog Learn About Our Cancer Collection Pages Plasmid Grant ...

...updates to iGluSnFR and SF-iGluSnFR have made it clear that it’s time to update our iGluSnFR post! Here...give researchers the ability to monitor glutamate release from neurons and other brain cells in vivo. Glutamate...protein binds to glutamate, conformational changes lead to an increase in fluorescence intensity from the...differentiating between synaptic vs. extrasynaptic glutamate release. Building on the previous generation of the SF-...Resources Additional Resources on the Addgene Blog Learn about other Fluorescent Biosensors A Practical Approach...

...than 75% photobleaching after just 1 minute of exposure. MG-2I-dL5** experienced minor bleaching after longer...ablate cells in culture and in vivo.  Read on to learn more about this killer illumination technique! ...real time.  In both cases, dL5** was tagged with mCerulean3 (mCer3) for visualization and expressed from ...Sign Up to Receive the Fluorescent Protein eBook Learn about Small Molecule Fluorophores Resources on ...

...incorporation of a 3’-5’ exonuclease in the reaction mixture. The exonuclease removes mismatched 3’ overhangs...guide the joining reaction. BioBrick Assembly will leave scar regions at the site of fragment joining, but...still creating scarless plasmid inserts.  Want to Learn More about DNA Assembly? Check Out Our Plasmids ...      Additional Resources on the Addgene Blog Learn More about Gateway Cloning Gibson Assembly and Other...

... of the mRNA, leaving only a red FP bound to the reporter. Because it contains a nuclear localization ...cell. When cellular mRNAs are digested with ribonucleases, ribosomes protect the segments of mRNA to which...occur. For a TRICK reporter mRNA this displacement leads to a change in the fluorescent signal: untranslated...Addgene. Additional Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background...

...libraries Learn about other members of the Addgene Advisory Board Additional Resources Learn a little...post-doc, Doench worked on using RNAi screens to learn more about cancer biology in Ed Harlow’s group at...and “with CRISPR, just the results are so much cleaner. The time frame from doing your primary screen ...Institute here Read other blog posts about CRISPR Learn more about the Broad Institute and the Platform ...

...ride is a great activity for both exercise and clearing your mind. And because the outdoors is so huge...events. Download Addgene’s Science Career guide to learn more about career paths and things you can do to...Plan out meals and snacks ahead of time.  Find learning activities for kids. For example, Skype a Scientist...collected into figures for papers or presentations. Learn a new skill for analyzing and visualizing your data...introductions and literature reviews could make the months leading up to your defense less stressful. Think about...

...complex processes can’t be overstated, we can also learn a lot from Nature’s simpler solutions to transmitting..., and they can rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS. For a more...appealing target for bioengineering, but it wasn’t clear whether they could 1) be decorated with proteins...Tunable Protein Piston That Breaks Membranes to Release Encapsulated Cargo. ACS Synth. Biol. (2016). doi...

...a team that created a novel form of AI that can learn new tasks and beat humans at the ancient Chinese... side you’re sure to have many opportunities to learn about and develop technologies that will have impacts...pulled tools from across the biological spectrum to learn more about the brain and neurological disease. I...better understand how aberrations in brain biology lead to altered function or maybe your favorite FRET-...pathways. Many scientists came to the Addgene booth to learn more about these tools and to tell us how useful...

...the following information for a Cas enzyme: Nuclease properties (CasID): Cis/trans activity, PAM requirement... (one of the most frequently used Cas enzymes) cleaves both strands of its dsDNA target in cis and has...generates staggered DNA nicks at its target in cis and cleaves ssDNA in trans. In sum, CasPEDIA documents the ...along with its action on the substrate (nick, bind, cleave, mixed). This information represents the ‘big picture...Perhaps you are a structure junky who just wants to learn about Cas domains. CasPEDIA either directly has ...

...illumination. This ability to reversibly bind and release a protein of interest in response to light allows...reaches the nucleus where gene expression changes lead to cell proliferation and differentiation. The lab...739201.  Additional resources on the Addgene blog Learn about using nanobodies to control protein activity...Read about RANbodies, reporter nanobody fusions Learn more about using the secondary nanobody toolbox ...Addgene.org Find nanobody expression plasmids here Learn more about optogenetics in Addgene's optogenetics...

...inspired you to perform this analysis. Having a clearly stated, specific goal for each analysis will help... currently working in) every time you use them, leave them in their own folder. Doing so will ensure that...same directory where it was run. This technique leaves a trail that further helps you keep track of your... entered is useful for beginners who are still learning the basics. However, these notes take up a lot...Read Our Tips to Keep Your Lab Notebook Organized Learn about Software You Can Use to Save Time and Track...

...Addgene’s blog to learn about making your own competent cells and our protocols page to learn about bacterial...have studied how HGT occurs in nature and have learned how to introduce genetic materials into cells in... and can include plasmid DNA or fragmented DNA released into the environment. Various factors promote ...more viral particles. Eventually the cell lyses releasing the newly formed viral particles that can infect...

... a date or two for beer, coffee or meals with colleagues you know will attend the conference. Invite others...sure your name tag is always visible. Some people learn names by seeing them. It is OK to ask someone to...the same talk. Read more on this advice to “never leave an empty seat” here. If there are communal meals...especially if they are not in academia. Make sure to learn how to use LinkedIn properly. Then, follow LinkedIn... lunch twice turns a stranger into a friendly colleague. Recommended Reading & More: Scientist Networking...