...very little data on which might work in your application. It can be quite difficult to decide on which...be searched and sorted by gene/target, name, application, pass/fail rating, sample species, antibody species... how the antibody performed in the submitted application, along with a brief description of the experiment...experiment. If that data was included in a publication, you'll find that information in the report as well... section will vary slightly depending on the application — flow cytometry, western blot, immunohistochemistry...data showing how the antibody worked in your application. The submission process is easy, with simple,...

...window onto communication between the muscle and brain, which may have therapeutic implications, say Hyo ...metabolites that are dynamically secreted by muscle during exercise to meet the brain’s physiological demands,” ...protects the brain from depression. Mice that are genetically modified to produce extra PGC-1a1 only in their..., the brain is protected from stress-induced biochemical changes that could otherwise lead to depression...

...speak for you (and you for them) Communication Being able to communicate effectively is part of what makes...excellent Technical knowledge and creativity Become a “thought leader,” develop a technical “niche,” develop...productive. Take all the opportunities you can to be communicating about science with others. If opportunities ... (be on a speaker committee) Follow industry publications like FierceBiotech to “learn the business” Attend...

...clones, it’s wise to quickly assess whether a significant number of cells have been edited. For many experiments...enzyme screening for resistivity to digestion, indicating targeted edits.   Interested in this method...sequencing confirmation. All knock-ins A method applicable to knock-ins both large and small is a software...consider using a version of Cas9 that has been specifically engineered for reduced off target editing, such...Tao Chen, Jos Jonkers, Bas van Steensel. Easy quantification of template-directed CRISPR-Cas9 editing. (...and variant-aware in silico off-target site identification for CRISPR genome editing. (2019) Bioinformatics...Resources at Addgene: Read our CRISPRGuide Get practical help from our CRISPR Protocols Find gRNA Design...

...which to test the validity of your biological model and the technical performance of your assay. But, in... a more-typical scale of tissue culture while still offering the potential for true biological insight...components, the model system and the assay are just as critical for success. Sometimes, you might have a few positive...facilitating more rapid follow-up. We hope the biological community will find these reagents to be useful...

...al. "Systematic identification of signaling pathways with potential to confer anticancer drug resistance... therapies,” said Kris Wood of Duke University Medical Center. “While we do not yet have a comprehensive... of 17 signaling pathways that are frequently implicated in cancer cell proliferation, survival, differentiation... make the library usable for pooled screens. Applications of the cancer pathways ORFs kit Wood and his... he said. “It could [be used to screen for] morphological features, migration, or survival in different...

...Thousands of mutations have been implicated in cancer. Yet, the majority of them remain uncharacterized...annotate genetic perturbations that may play significant roles in cancer pathogenesis. Their work culminated... of mutations they identified included gene amplifications and fusions, missense mutations, and indels...mutations were found to be “neutral,” having no significant effect on cell viability compared to a wild-type...Approximately ~22% of the mutations, however, significantly increased cell viability, and these “activators...aberration drivers of pancreatic cancer." Nature communications 7 (2016): 10500. PubMed PMID: 26806015. PubMed...

...with the gRNA), application, cas9 species, PubMed ID, and depositor. The CRISPR applications we currently...shown to work in CRISPR experiments. Addgene’s dedication to resource sharing has led us to develop a searchable...validated gRNA Target Sequence Table. Disclaimer: The efficacy of gRNAs is impacted by the target location, target...the carats in the datatable headers to sort alphabetically or in ascending or descending order, in the...and final experimental outcome in the original publication. As you develop and confirm new gRNAs, please...

...Since the first research applications of GFP were published in the 1990s, biologists have spent a lot... mammalian cells, and it may find wide use in biological research. Photoconvertible fluorescent proteins...color was observed after excitation at 488 nm, indicating that spontaneous conversion did not occur. Tobacco... Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP photoconversion One concern...2):131-3.  PubMed. Find plasmids from this publication at Addgene. Subach OM, Patterson GH, Ting LM...nmeth.1664. PubMed. Find plasmids from this publication at Addgene.   Addgene’s Fluorescent Protein...

...scientific community support open sharing. But, one critical part was missing when the COVID-19 plasmids first...Carolina at Chapel Hill University of Illinois Chicago                     Boston Children's Hospital                           ...                           National Institute for Physiological Sciences Broad Institute                                       ...                                       Chinese Academy of Medical Sciences University of Texas at Austin University of California...part of research projects, another way is to systematically offer thousands of constructs for tagged and...enables research on solute carriers (SLCs) for medical research Recently, the Michael Davidson Fluorescent...

... QC/QA team as a DNA Sequencing Intern. The application process When I first saw the comprehensive job...immediately clear that Addgene wanted to ensure applicants understood what the expectations were for the...the intended position. After submitting my application, I was contacted to participate in a brief fifteen...the depositing labs intended. With the use of applications like SnapGene and Geneious, I was able to determine...team and shows how they fit into the system. Specifically as a co-op, it is encouraged to explore different...currently working on projects for the Science Communication function at Addgene. Working a 9-5 As it turned...

...technology, the Vale and Weissman labs took this biological lesson and created a system to amplify fluorescent...modified the GCN4 sequence to increase its alpha-helical structure and stability, creating v4 SunTag. Since...aggregation, it’s recommended for most imaging applications. What can you do with SunTag? Tanenbaum et al...flames,” it’s clear that the system is broadly applicable across many areas of research. Beyond fluorescence...activation of target genes, but we’ll save that application for another day! References 1. Tanenbaum, Marvin...et al. “A protein-tagging system for signal amplification in gene expression and fluorescence imaging....

...generally linked to dopamine (DA), a well-known reward chemical and neurotransmitter. But aside from DA’s famous...at a certain frequency, and this frequency can indicate behavioral state, for example, if someone is awake... it can be performed in a live awake animal by surgical implantation of the probe into a specific brain...in freely moving mice Record DA dynamics of optogenetically activated and inhibited neurons in live mice...neuronal imaging of dopamine dynamics with designed genetically encoded sensors." Science 360.6396 (2018): eaat4422... PMCID: PMC6287765. Sun, Fangmiao, et al. "A genetically encoded fluorescent sensor enables rapid and ...

...swimming where they periodically beat their tails left and right in a symmetrical motion to move forward...behavioral studies. In neuroscience, opsins are typically ion channels that allow charged particles to cross...Kolmer-Agduhr cells was sufficient to trigger the symmetrical tail movement unique to spontaneous slow forward... photostimulation in order to drive the full symmetrical response.  The function of Kolmer-Agduhr cells...Aristides B., Filippo Del Bene, and Herwig Baier. "Optical control of zebrafish behavior with halorhodopsin...

... (e.g. 6xHis containing pDEST-HisMBP). After purification the protein can be released from the resin and...types of yeast expression vectors. Consider your applications and experimental design. You can create a new...more traditional approach of an autonomously replicating vector that carries either a wild-type version...expression pattern of a defined promoter region, ectopically expressing YGOI in a pattern that allows the ...a variety of other types of vectors for most applications used to study gene function. From making transgenic...transgenic animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene ...

... Foundation Round Table. In any field, good communication skills help when it comes to finding a job. ...an argument for taking the time to hone your communication skills. So, where can a graduate student or ...opportunities than ever before for scientists to communicate about their work and their interest in science...your area or at your institution; the news or communications office is a good place to start. It’s not unusual...about science writing as a career, the Science Communication Program at the University of California, Santa... your research stories. Consider practicing communicating your research by guest blogging here, just leave...

...problems, develop general technical proficiency, communicate your work, maintain ethical standards and interact...? Listen to our podcast interview with Harvard Medical School researcher Connie Cepko to learn about her... you admire her management style? Does he have ethical behavior? Does the lab encourage diversity? Knowing...websites listing alumi and it is easy to use publication records, university websites, ResearchGate and...

...appreciable levels of modification. Whole-genome sequencing of cells modified by CRISPR indicates that the consequences...20nt-NGG] and [5’-CCN-20nt], as it is equally efficacious to target the coding or non-coding strand of ...desired location of edit; efficiency decreases dramatically when the cut site is >30nt from the proximal... via CRISPR cutting. For CRISPRa, it is most-efficacious to target a ~100nt window upstream of the transcription...sequence may not be available. Predicting gRNA efficacy We and others have examined the ability to use...needs to test in the lab. Importantly, for any modification of interest, it would be unwise to make conclusions...

...natural and engineered repressible promoters. Chemically repressible promoters Tet-Off In its native context... GAL80 under different promoters and create sophisticated patterns of UAS-driven gene expression. To further... PMCID: PMC2942869. Find plasmids from this publication at Addgene. del Valle Rodriguez, Alberto, Dominic... PMCID: PMC2883883. Find plasmids from this publication at Addgene. Wei, Xing, Christopher J. Potter... PMCID: PMC3846601. Find plasmids from this publication at Addgene. Riabinina, et al. “Improved and ... PMCID: PMC4344399. Find plasmids from this publication at Addgene. Riabinina O, Potter CJ. The Q-System...expression of genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS Yeast Res...

...supercoiled DNA ends to facilitate replication. The TOPO technique specifically uses Vaccinia virus-isolated...insert comes from using Taq polymerase for the amplification step since Taq polymerase leaves a single deoxyadenosine...minutes (incubating the full 30 minutes will not significantly improve the transformation efficiency).  5. ... has an error rate of about 1 in 3,500 bases. Typically polymerases with proofreading functionality are...are planning on transforming your reaction into chemically or electro-competent E. coli - excess salt causes...