... information you are looking for in our catalog or educational resources: plasmid, pooled library, blog...articles. From paper to plasmid Authors regularly indicate that their plasmids are available from Addgene...papers. Maybe they started their deposit prior to publication and were able to include specific plasmid ID ...terms we can use to find materials from this publication. Searching for the title of a paper works You...filters Not every plasmid search will start from a publication. You may have an idea for a plasmid that would...try different terms you may find the available categories and filters change as we now only show options... a second filter, this one from the ‘Requests’ category, will leave us with a single plasmid to consider...

...about which reagents were used? Unambiguous identification of a reagent is crucial for reproducibility...can lead to wasted time or retractions. Using a catalog number for identifying a reagent can be helpful...reagent online. Furthermore, if the link between the catalog number and reagent description is lost (e.g. a ... supplier is bought out by another company and catalog number changes) the original identifier loses its... but this approach has been adapted for the identification of physical materials that we use in our labs... which serve as unique PIDs within the Addgene catalog. These IDs allow you to unmistakably identify a...item. Given a specific Addgene ID # listed in a publication, you can quickly pull up the associated plasmid...

...Few Complicated Non-natural amino acids 1 Various Many Complicated   Applications Organic...new green fluorescent protein, some affinity purification tags, another version with that same red fluorescent... an enzyme that has been mutated to block its catalytic activity and instead become trapped in a covalent...having to clone a whole new construct for every application. Plus, the photophysical properties of chemical...advantages over fluorescent proteins, opening up new applications.   Figure 1: Live-cell labeling strategies...Although these multi-component systems can be complicated to implement, they can be highly specific with...self-labeling tag and a fluorescent protein) enables applications like multi-color imaging (Figure 2). If you’...

... post on mammalian vectors, will catch you up on the core replication and resistance features of yeast...recombination. Yeast Replicating plasmids (YRp):  These vectors contain an Autonomously Replicating Sequence (ARS...vectors must contain a yeast-specific origin of replication (ORI) and a means of selection in yeast cells...antibiotic selection markers.  The yeast origin of replication (Please note: This first section primarily pertains...ve also noted some features required for the replication of fission yeast, Schizosaccharomyces pombe, ...within a yeast vector determine how the plasmid is replicated and maintained within the yeast cell. These elements...integrated into the host DNA or is independently replicated as an episome. The four main types of yeast plasmids...

... you with two measurements: forward scatter (FSC) and side scatter (SSC). To acquire these measurements...exclude all duplicate events (two cells stuck together) from your single cells, as duplicates can’t be reliably...types of scattering that occur when a cell is pulsed by the laser. Within each type of scattering, you also...to keep it short though, promise! FSC and SSC: scattering To digitally extract your cells based on their...passing cell with a laser and detects the light scattered by that cell at a low angle (FSC) or large angle...: The laser pulse illuminates the cell and is scattered forward at a low angle (FSC) or to the side at...passes through the laser beam and causes it to scatter (FSC/SSC), the detected photons can be represented...

...sister locations on April 22, 2017, scientists and non-scientists alike will march to advocate for science...already come to pass. It may only be by advocating for science education and literacy that this changes.  2...marching for. I will be marching to advocate for science education and literacy as I believe people that...communities, microbial interactions, and science education.  Disclaimer: The views presented in this article...science’s place in education, government, and civilization in general (1). Science and politics? Science...process begins with a testable hypothesis - an educated guess about how some part of the world functions...health Immigration Pollution Healthcare Science education To make scientists relatable (17, 20). This...

...sensors One of the latest additions to Addgene’s catalog is a new class of sensors for neuropeptides and...doi: 10.1038/s41592-022-01597-x. New glutamate indicator iGluSnFR3 Improving on the glutamate sensor iGluSnFR...They generated variants optimized for different applications (v82 has higher affinity for glutamate but slower...PDGFR.codonopt    Aggarwal A, et al. Glutamate indicators with improved activation kinetics and localization...have developed a new genetically-encoded voltage indicator (GEVI) called JEDI-2P (Liu et al. 2022) optimized...Sustained deep-tissue voltage recording using a fast indicator evolved for two-photon microscopy. Cell. 2022 ...has generated a new genetically encoded calcium indicator using small monomeric NIR fluorescent proteins...

...production and purification that yield high titers (see also AAV protocols for production, purification and titration... are replication incompetent and cannot readily be passed on to subsequent cells via replication like ...intracerebral application, which is more invasive and has a much higher risk of complications. References...between different brain regions. For example, communication between the hippocampus and the prefrontal cortex...intracellularly spread away from the site of application and aid in visualization of the morphology of...flow in order to understand which brain regions communicate with each other and how they do it (i.e. where...where the signals come from and what implications their signaling is likely to have). Anterograde tracing...

...Restriction Enzymes + - PCR Amplification + + Digestion + + Gel Purification + - Ligation + - Transformation...vector is located in the 3’ end of the multi-cloning site, and the reverse primer is located in the 5’...Molecular cloning is the isolation, insertion and amplification of a recombinant DNA without sequence alteration... multi-step process consisting of digestion, purification, ligation, and transformation. While restriction...sequences for base pair overhangs, insert and vector purification, and, most importantly, excessive time requirements...is ligation independent, it does not require purification of insert or vector products, nor does it require...digested mixture. There is no need for further purification before transformation. After transforming cells...

...estimated scatterplot smoothing. (C) Number of requests per year for pooled libraries, categorized by function...insertion via a chemical modifier. We used these categories to see what kinds of CRISPR tools people were... pooled libraries, although we used different categories (Figure 2C). Note that in Figure 2, the numbers...Cumulative number of deposited CRISPR items, categorized by CRISPR function, by year. (B) Number of requests... years.  Libraries are divided into different categories based on function: knockout (KO), activate, inhibit...Figure 3, which shows requested CRISPR plasmids categorized by “age” or year they were deposited, you can...Percentage of requests per year for CRISPR plasmids categorized by year of deposit. Outlines show a representative...

...addgene.org if you would like to contribute to the catalog. The Catalog Will Start With Neuroscience Antibodies ...performance in various applications are included in the full webpage. Educational Resources As part of...resource, which includes recombinant antibodies, educational materials, protocols, and shared data, on our...sequence information for the antibodies in our catalog, so that you will know exactly what you are working...community about how the antibodies work in various applications, enabling you to better plan your experiments... improving research infrastructure, from our educational materials to our repository of open samples. ...your research, Addgene is creating an array of educational materials to accompany our antibody collection...

...Effective verbal and written communication skills are key to publication and acquisition of funding. If...  Writing Communication Publicity Consulting internships Communication Teamwork Management...Models Communication Regulatory knowledge Networking Blogging Writing Communication Networking...scientists identify potential career pathways, education and mastering new skill sets must occur by doing... skills such as leadership, management, and communication. A great approach to fully understand and acquire...find open positions and view their required qualifications and skill sets Taking the time to identify... science class: Science Club for Girls) Communication to diverse audiences Organization Mentorship...

...or DNA base pair mismatches (8-11). For some applications, there cannot be any end-joining activity, and...regions. Why use a high fidelity ligase? Numerous applications have been developed to take advantage of the...Taq and other thermostable DNA ligases. These applications detect specific nucleotide sequences with high...rounds of melting and annealing allows linear amplification of the probe ligation product. With the right...uses two sets of probes to achieve exponential amplification. What does fidelity mean for a ligase? DNA ligases...conditions to be screened per plate. The results indicated that the optimal buffer for Taq DNA Ligase contains...Ligase are not replacements for T4 DNA Ligase in applications such as routine cloning or DNA library preparation...

...in 2018. By the end of 2023, our catalog had grown to 879 viral catalog items (Figure 1). The service caught...repository. GCaMP, a calcium indicator (biosensor, by Addgene’s categorization) was first developed in 2001....   Figure 2: Numbers of AAV tools, by category, available from Addgene as of 2023. A large... calcium and voltage sensors, are the broadest category of viral vector tools, which may help explain ...number of requests for vectors in the “controls” category, which includes things like viral vectors that...capsid of the AAV, many of the vectors in Addgene’s catalog are offered in different serotypes. In fact, the...are the most requested serotypes (Figure 4), our catalog includes vectors in a variety of serotypes, including...

...mice. B) Quantifications showing high microglia specificity of GFP expression for the indicated ssAAV5s...ssAAV5s. C) Quantifications showing high microglia transduction efficiency for the indicated ssAAV5s. Figure...3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral integration, producing a second...described in Anzalone et al., 2019, including modifications to the Cas9 or reverse transcriptase enzymes...enzymes, inhibition of host cell proteins, and modifications to the pegRNA. The Britt Adamson Lab recently ... vector preps expressing the Voltron2 voltage indicator, created by the Schreiter Lab and the GENIE Project... available! Building on their earlier Voltron indicators (a fusion of Ace2N and HaloTag, Figure 4), they...

...also a science communicator, UBC Public Scholar, and a member of the Vancouver SciCATs. Follow her on ...researchers and supporters of science marched to advocate for scientist participation in government, evidence...promote help motivate you to spend a little time advocating for science and getting yourself out there for...April because devaluing science has alarming implications for our public policies—many of which impact...scientist. I’ve worked tirelessly to get the publications, grant funding, and teaching experience that...As scientists we spend years being taught to communicate in a very specific, technical manner, so much...advocacy initiative that seeks to bridge the communication gap between scientists and the public. Find ...

...molecular biology applications for biotin and avidin. A short list of some other common applications and links...downstream application. Biotin labelled oligonucleotides can be used in a multitude of applications including... carboxylase and decarboxylases. These enzymes catalyze the transfer of carboxyl groups to organic acids...this highly specific post-translational protein modification. Most organisms have less than five enzymes ...uses of biotinylation in the lab are protein purification and immunoprecipitation experiments. Biotinylated...sites found on avidin are also ideal for signal amplification during these experiments. During detection, ...bound to your protein of interest leading to amplification of the reporter signal and easier detection....

...proteins within a system. This can be pretty complicated given that a single cell has tens of thousands...single protein? Oftentimes, they rely on protein purification. In this strategy a single protein can be isolated...this blog will focus on one of the most common, purification based on the polyhistidine tag. What is the ...purity (Janknecht 1991, Hochuli 1988). In this purification approach, the metal ions are immobilized on ... incorporation of a cleavage site and a post-purification cleavage step to remove the tag in order to ...2000).  One downside of polyhistidine tag-based purification, however, is the propensity for nonspecific ... there is no cut or dried answer. Successful purification requires the tag to be accessible to the metal...

...the OpenWorm website. microPublication microPublication is a unique publication for the worm community...into larger publications and would otherwise be lost in lab notebooks forever. microPublications are peer-reviewed...aims and approach of CeNGEN, check out their publication.   3’UTRome The 3’UTRs of mRNA are important ...normally one figure and can be negative and/or even replicated studies to show reproducible results. This a ...referenced. In addition the data published in microPublications is curated and added to other databases including...including wormbase and flybase. microPublication is run and maintained by members of the worm community...introducing point mutations that was published in a microPublication (Schwartz and Jorgensen, 2018).  These are...

...post-translational modifications, quantify protein levels relative to standards, detect the cellular location of proteins...pull-down assays.  Westerns are divided into two categories, native and denaturing. In a native western, ...matrix, the blotting membrane, the gel, a copper cathode/iblot gel matrix, and filter paper. Pretty much...search, first pay attention to the validated applications listed on the vendor’s website and choose an...western and vice versa.  In addition to validated applications, pay close attention to the controls listed ...and incubation times. If the vendor includes publications that cited the antibody, consider reviewing ... the materials and methods sections of those publications.  Finally, always verify that the chosen antibody...