...ChIP-seq methods provide a wealth of information about how protein networks work in cells. A major bottleneck...two-part cr/tracrRNAs for CRISPR experiments (in collaboration with Integrated DNA Technologies, USA). These...commercial suppliers without investing time and labour on reagent preparation in the lab. Figure 3: csRNP-assisted...first crRNA we designed for ~250 genes, we have had about 30% tagging success, although many of these genes... When not pipetting, he can be seen daydreaming about sunny days in the UK.   References 1. Bressan RB...

...post, a study coming out of MITs Human Dynamics Laboratory identified five characteristics of very successful...conversations within the team. We are not talking about gossip here – this means encouraging a culture of...Medical School researcher Connie Cepko to learn about her management style.   Seeking feedback frequently...information to make change whenever possible. Be public about changes influenced by feedback. If you don't do ...

...Joanne Kamens, Melina Fan, and Ken Fan each spoke about what Addgene means to them. Ken mentioned Addgene...Addgene could not do what it does.  Melina talked about her grandfather’s humble beginnings as an operator...and add to this post. Interested in learning more about the last 10 years @Addgene Check out our impressive...stats from 10 Years of Plasmid Sharing Or read more about what happens Inside Addgene   ...

...the Addgene blog Learn more about Addgene's new search tool Read more about other features of Addgene's.... Instead of trying to place all of the content above the fold, we decluttered the page elements and allowed...comfortably tell our story, to get scientists excited about what Addgene has to offer, and to help scientists...

...not even get a solution to the problem you called about. Next, you try an email, given that this is the ...” next to an encircled question mark (see image above). This useful widget allows you to access our Help... or you just have a general question or concern about the answers we provide, within the widget you also...Addgene Blog Read other Inside Addgene Posts Learn about New Features on Our Plasmid Pages Resources at ...

...blog Learn about Gateway cloning Read about genome engineering in flies Learn more about controlling ...the same genetic makeup, overcomes all of these labor-intensive and invasive techniques to reliably label...

...attended the Go green! Innovative practices for laboratory waste symposium, sponsored by New England Biolabs...Biolabs and Labconscious. I was eager to learn about practices I could bring both to Addgene, and share...offsets the cost of regular waste services. Learn about how to start a glove recycling program at your institute...lab tips for your research Read another blog post about conservation in the lab Resources on Addgene.org...

...We’ve talked a lot about the quality control process at Addgene by introducing our new sequencing partner...partner seqWell and going into detail about how we use next generation sequencing results to perform quality...control on deposited plasmids. We’ve also talked about how our new Snapgene generated maps provide improved...open reading frames, which is often our first clue about where the gene of interest might be located. By ...

...sensors generally follow the same pattern described above - a sensing domain induces changes in a fluorescent...expressing a biosensor in eukaryotic cells, think about the best place for the biosensor to be expressed...MinDis).iGluSnFR. Picking your reporter Don’t forget about the other end of the biosensor: which fluorescent...Blog Check out the Rosella Autophagy Sensor Read about Aptamer Fluorophores Use CRISPRainbow for Genome...

...successful writer. When I originally talked to my PI about becoming a better writer, I was unsure if I should... a manuscript and critiquing it, were not too laborious, but helped build my understanding of how to generate...and the writing workshop. SWPI's writing workshop About a month later I received another email notifying... manuscript, scientists have a tendency to be elaborate and over explain the material. The point of writing...

... for the Addgene blog, I was focused on writing about new scientific techniques and cool plasmids. Creating... graphics were usually the last thing I thought about when writing posts. Since then I’ve realized my ...know of a tool or resource not mentioned? Tell us about it in the comments section!  Additional resources...scientists Find science career blog posts Learn about other lab software ...

...approached me and asked about my current projects and personal interests. We chatted about social media analytics...while eating lunch. There was something so special about her pure interest and curiosity to understand my...hours! As my time here comes to an end, my worries about making connections with coworkers seem, in retrospect...

...scientists, including Nobel Laureates, who speak about their work and current, trending topics in their... The society facilitates opportunities to learn about Synthetic Biology and exciting new technologies ...opportunities for society members as well as arranging collaborations and conferences for their students. Another...interests. Scientific societies like those mentioned above provide strong foundations upon which young scientists...

...research question that people (especially you!) care about. Have you critically evaluated your plan? Have... to complete and has multiple benchmarks. Think about what your personal goals are and set milestones .... Take the time to generate new ideas and think about them critically before diving in. Research projects...Medical School) for taking the time to speak with us about adding new scientists to their labs. Read All the...

...nascent heteroduplex and abolish cleavage (Pattanayak et al., 2013). As mentioned above, however, Cas9 sometimes..., 2022), indicating that there is more to learn about this process!      Figure 3: Crystal structure...rest of Cas9 and is highly flexible; it can rotate about 140 degrees between inactive and active conformations...conformational changes that make Cas9 such a valuable laboratory tool are still an area of active research. New...

...science,” she says. In 11th grade she heard a talk about Z DNA that inspired her to major in molecular biology...was through this connection that she learned more about Addgene. “The mission to share critical reagents... molecular research, but also became passionate about virology,” he says. He was working in an HIV lab...gene therapy either through direct industry collaborations, grants, or setting up commercial and nonprofit...

...Addgene blog Learn about other Cas proteins Read all of our CRISPR blog posts Read about GeoCas9, another... able to eliminate the need for axenic (pure) laboratory cultures to study these microbes. From 16S rRNA...by flexibly deploying IgnaviCas9 without concern about its stability under harsh conditions. One example...

... Addgene blog Read blog posts about synthetic biology Learn more about codon usage biases Resources ...scientists from MIT tapped into this data to learn about trends in synthetic biology and DNA synthesis. They...synthetic just by looking at its sequence. MIT collaboration for gene classification This inquiry started...

...flexibility needed for multiplexing. In the case above, you can’t costain with your desired antibodies ...multiplexing with chimeric antibodies. In the case above you should use an anti-rat IgG1 secondary antibody...this blog post has answered all of your questions about the utility of isotype conversions and chimeric ..., Bright, S. M., Robinson, M. K., Graham, E. T., Sabo, J. P., Owens, R., King, D. J., Norris, S. H., &...

... with the three other organizers. We asked them about their experiences as they navigate the new norm ...pre-workshop questionnaire to the participants to learn about what they were looking to learn in the workshop ...having a stable internet connection,” says Abitua about her workshop. “We made sure that we had a backup...on the Addgene blog Read all of our blog posts about conferences Find 25 tips for working from home ...

... to find that there is insufficient information about which reagents were used? Unambiguous identification...available through Addgene and find verified information about each plasmid item. Given a specific Addgene ID #...the Addgene website and find specific information about the plasmid: name, description, how the plasmid ...out this post on Scientific Reproducibility Learn about the anatomy of a plasmid page at Addgene Additional...

... in culture and in vivo.  Read on to learn more about this killer illumination technique! What is a photosensitizer...within the near-infrared (NIR) spectrum (>620 nm). Above is a brief schematic of how the MG-2I-dL5** FAP-...use MG-2I-dL5** with your research? Let us know about your experiences using photosensitizers in the comments... to Receive the Fluorescent Protein eBook Learn about Small Molecule Fluorophores Resources on Addgene.org...

...saved and cannot be deleted, so there is no worry about misplacing your results or losing that lab notebook...enzyme, or name to discover all the information about that restriction enzyme or share it via email with... this app. However, if you get quizzed randomly about a hazardous chemical and can’t find your lab’s MSDS... new researchers or students beginning to learn about biochemistry. The least this app will do is allow...

...posts on base editing Read more about CRISPR Learn all the basics about CRISPR in our CRISPR 101 blog series...both changes simultaneously with 54% efficiency, about 7 times higher than ABE7.10. However, the increased...change C to T, G to A, A to G, and T to C, represent about two-thirds of known pathogenic point mutations. ...

...Addgene blog Learn about using nanobodies to control protein activity Read about RANbodies, reporter ...reporter nanobody fusions Learn more about using the secondary nanobody toolbox for immunodetection Resources...Find nanobody expression plasmids here Learn more about optogenetics in Addgene's optogenetics guide ...

...teachers in the U.S. and beyond. “I want to learn more about Synthetic Biology. Can you tell me where to start... for scientific exploration, discovery, and collaboration. One module currently under development comes...circuits and logic gates. This BioBuilder-Caltech collaboration has resulted in a new BioBuilder book chapter... or small, receive the BioBuilder textbook, a laboratory kit, and support during the academic year from...

...Since their first publication in 2012, the Nagai laboratory has assembled a collection of multicolored nano-lanterns...Nano-lanterns, Takai et al. continuously imaged cells for about 4 hours! The latest enhanced Nano-lanterns (eNL)... Nano-lanterns and your experiments The Nagai laboratory has a done a wonderful job characterizing Nano-lanterns...Additional Resources on the Addgene Blog Learn about Photoactivatable Fluorescent Proteins Check Out ...

...the name, date, and location of all analyses (see above). Give every experiment its own directory: In ...record your goal, approach, and conclusions (see above). Keep original files in a separate folder: You... can simply make a note in your methods section about the edit that was temporarily used. Just be sure... Tips to Keep Your Lab Notebook Organized Learn about Software You Can Use to Save Time and Track Your...

...Leah Bury of Microscopic Art. I started thinking about this problem when I recently attended the 2018 Chromosome...lot of scientists who not only remember to think about DNA in this context, but also specifically study...plasmids containing these genes. As with the PIs above, many of these genes were discussed at the 2018 ...chromatin This is not the first Addgene blog post about chromatin--multiple Addgene colleagues and guest...

...species (read Arbor Biotechnologies' press release about their Cas13d deposit here). Konermann et al.: CasRx...this paper are available at Addgene. Learn more about research at Arbor Biotechnologies. Additional Resources... Resources on the Addgene Blog: Learn about CRISPR RNA Editing with REPAIR Get tips for using Cas9 Transcriptional...Transcriptional Activators Learn about Cas13a (C2c2) Characterization Resources on Addgene.org: Find...

... Get Started with AAV Learn more about Lentivirus Find out More about Addgene's Viral Service Resources...We often think about the brain’s function in terms of its regions. But equally important is the way the...head bone’s connected to the neck bone,” but with about a hundred billion neurons and six hundred trillion...

...applications to help you start (safely!) thinking about the possibilities. Ready to plan your next (or maybe...S, Varmus H. Retroviruses. Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY, USA: 1997. PubMed...Additional Resources on the Addgene Blog Learn More about Lentivirus Listen to Connie Cepko Discuss Gene Therapy...Addgene.org Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors...

... called NABOR (Neuroscience AntiBody Open Resource). The antibodies distributed through NABOR are recombinant...troubleshooting methodologies drew me to learn more about the process that was used to create this important...secondary antibodies are commonly used in research laboratories to detect the different subclasses, but there...

...from previously discussed methods. As described above, you design two primers, each containing the desired...suited for insertions and deletions. The figure above shows guidelines for primer design; a desired insertion...mutation-containing colonies/total colonies) is about 84% for each of the three methods. However, REPLACR-mutagenesis... Additional Resources on the Addgene Blog Read about Other Plasmid Cloning Techniques like SLIC and Golden...

...antibiotic selection here at Plasmids 101, we can talk about an even more specific method of screening your cloning... plasmid is a great start when cloning, but how about being able to choose those that contain plasmid ... a widely used method to do just that! (To read about other screening strategies, check out our screening... called α-complementation. The system described above was put to practical use in the following way. Scientists...

...that screen with a system you are really confident about working for most genes and individual targeting ... system, Konermann explained, was new knowledge about Cas9’s crystal structure. “I thought maybe we could...RNA we use. What we have to do now is learn more about the rules.” He said that there is no theoretical...References & Further Reading: For more information about SAM, including a tool for getting optimized guide...

...transitioned away from the bench and became all about finding the best product-market fit for this set...Notably, the team must have an upfront conversation about its goals and vision for the company; this will ...section! And read on for more great information about starting a biotech company. Suggested reading The...of Sample6) for taking the time to speak with us about the biotech startup process and work environment...

...fragment are we talking about here? Many of the Fab fragments described above can be used for similar ...fragments are produced by proteolytic digestion above the hinge-region, which results in two independent...you consider which tools to use, be sure to think about what pieces of the antibody you do and don’t need...

...August 2nd in Nature gives us even more to talk about, as it represents another step towards CRISPR germline...impossible with current tools, thus quieting some fears about the creation of “designer babies.” The differences...of individuals with varied backgrounds. Talking about this study with non-biologist friends Like me, you... Blog Check out Our New CRISPR Topic Page Read about High Fidelity CRISPR Variants Target Multiple Genes...

.... Gendered Innovations is an international, collaborative project—funded by the European Commission, the...research.  In addition to the studies referenced above, another excellent resource for research design ... whole human population. To learn a little more about where differences in effects on the sexes come from...cost. Especially in animal research, they worry about needing more study subjects and control groups. ...

...source of monoclonal antibodies, but they are laborious to generate and costly to share. What is a RANbody... particular applications. Read on to learn more about each reporter or check out table 1 for a summary...Additional Resources on the Addgene Blog Learn more about using the secondary nanobody toolbox for immunodetection...can be found here Watch this video to learn more about nanobodies Additional Resources Looking for primary...

...cloning before you can start your experiment? What about all the reagents you need to order? Sometimes, it...dive deep into generating the data you’re excited about. When we were closing in on the one million plasmids...million plasmids shared than not!), we think it’s about time we updated this post. How much time does ...choose the right cloning method Find blog posts about scientific sharing here Download the Plasmids 101...

...meetings, Twitter, coffee chats with potential collaborators, blogging and more. These each have different...key routes for researchers to share information about the emerging SARS-CoV-2 virus quickly and openly...awareness of the benefits of preprints. Some worry about faulty work being posted as preprints, assuming ...submission. Preprints at Addgene Preprints are all about sharing scientific knowledge faster and more openly...

... blog Read more about the PHP.eB AAV capsid Read more about optogenetics Learn about the differences ...library were designed using structural information about parental channelrhodopsins to create recombinant...

...One of the best things about sharing plasmids through Addgene is that we provide an added level of confidence...bp of clean sequence from a typical reaction, or about 250 amino acids worth, so make sure the feature ...96-well microplates (identical copies, see image above). We then select forward and reverse primers for...specifically to target this region, as outlined above. Avoid primers that bind within or must read through...

...Nielson and Voigt (2014).   We talked with Voigt about this collision that’s taking place between CRISPR...straightforward surprisingly. So I think a nice thing about CRISPR is how much easier it is to work with than...can slow their growth. Something has to be done about that.  The second problem is fundamental. We built...worked in each case. Sometimes in CRISPR people talk about off-target interactions. That’s relevant if you ...

...a broad range of reproductive, developmental, metabolic, and immune response programs. NRs also encompass...gallstone disease, senescent illnesses, and the metabolic syndrome. Coregulators Coregulators constitute... regulatory regime (such as genes involved in metabolism, growth or cytokine action). Primary or core ...and the free exchange of these vectors between laboratories. The development of the transient transfection...

...CRISPR Featured Topic Page Learn more about Cpf1 Find out about CRISPR Multiplexing Resources on Addgene.org...coli and other bacterial species. Read on to learn about the tools available for bacteria and some of the...that can be used in this way. As described in the above CRISPRi section, the Marraffini Lab published a ...as a collection of organisms, their genes, and metabolic processes, occupying the gut. Many human diseases...

...you do to get ready for this interview? Maybe the above scenario sounds familiar to you. It was my experience...positions often required a one-hour presentation about a prior research project. In total, the interviews... find lists that have lots of general questions about job responsibilities, performance evaluations, professional...require. Frequently they will ask you to present about your prior research. This is a chance to highlight...

...can be done using FRET! Read on to find out more about this amazing imaging technique and find further ...our curated list of biosensors too, as another laboratory may have already created the sensor that you ...ability of labs to answer sophisticated questions about cellular processes. Currently, FRET experiments ... Tips for using FRET in Your Experiments Learn about Fluorophore Targeting for FRET Studies FRET Based...