...Description Origin of Replication (ORI) DNA sequence which allows initiation of replication within a plasmid...point for PCR amplification or sequencing. Primers can be exploited for sequence verification of plasmids...important. Our “Plasmids 101” series designed to educate all levels of scientists and plasmid lovers - serves...plasmids are small circular pieces of DNA that replicate independently from the host's chromosomal DNA....virulence. All natural plasmids contain an origin of replication (which controls the host range and copy number...Minimally, lab-created plasmids have an origin of replication, selection marker, and cloning site. The ease... plasmids and the ability of plasmids to self-replicate within a cell make them attractive tools for the...

...of complex post-translational modifications. Extraction and purification of recombinant proteins from ...modified via an array of post-translational modifications including protein folding, formation of disulfide...mRNA and the addition of post-translational modifications Learn more molecular biology in our Plasmids... experimental, biotechnological, and medical applications. Researches can visualize proteins in vivo by...expression systems used for protein production and purification. These include mammalian, insect, bacterial,...separated from contaminants using a variety of purification methods. Deciding which expression system to...protein you need   Your plans for downstream applications In this blog post we will summarize some of...

...results.  Replicates Replicates are another critical control in biological experiments. Replicates can be...of biological replicates such as multiple mice in each group and C, technical replicates or repeated measurements...secondary antibodies.   Application-specific controls Some antibody-based applications have very specific ...then moving on to controls for antibody-related applications. After reading, you will have a better understanding...blot and look for the molecular weight shifts indicative of tau phosphorylation. By including the positive...statistical power of an experiment. Technical replicates are repeated measurements of the same sample ...difficult to obtain meaningful results.  Biological replicates are distinct samples that are treated identically...

...Science Communication Conversation in Science Communication The Purpose of Science Communication #1 Mistake...supporters of science communication. To help you think about ways to effectively communicate your science, we...News at Harvard University. Effective science communication is required to promote public support for research... organization that works to promote science communication. Science in the News hosts a podcast, seminars...That’s right, #ActualLivingScientists, eager to communicate their science. In this first half of our two-...programs SITN runs and learn how they think about communicating science. Listen to the interview with Science...Mistake Scientists Make when Communicating with Non-Scientists How to Approach Different Audiences Listen...

... 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...

...development, certification, degrees, foundations, coaching and even Twitter Chats dedicated to the Grant...the candidate application process, the candidate’s ability to write a grant application through a written... much more than writing. In fact, many grant applications and grant makers have moved away from long written...Now they require grant seekers to use online applications with significant character limits for the written...written portion of the grant application. They have increased their demand for stronger budgets with clear...duties can range from coordinating the grant application and submitting it, to evaluating the capacity...asked to serve on review panels and to review education materials for publishers. When the community orchestra...

...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...

...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...

... 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...

...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...

...mismatches on modification by AsCpf1 (red) or LbCpf1 (blue). Mismatch positions are indicated below each ... this publication at Addgene. Makarova, Kira S., et al. "An updated evolutionary classification of CRISPR-Cas...it lacks Cas9’s other HNH endonuclease domain, indicating that Cpf1 functions differently. Since Cpf1 loci...differing from that of Cas9 (3’-NGG) in both genomic location and GC-content. After sequencing and searching... off-target editing rates, determined using sophisticated genome-wide analysis. At many of its computationally...compared the ratio of total off-target to on-target modification for AsCpf1 and LbCpf1, and found that both orthologs...-target editing in a cell culture model. The application of Cpf1 to genome editing is exciting both in...

...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...