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...equipment sharing or a collaboration – it requires re-location of the scientist to the new institute where their...equipment! Preparing for your visit   Design and communicate a research plan Your time at the new research... and you are set up for success. If you are communicating with a professor or institute, ask if you will...out to your bench mentor (if you have one) to communicate your plan as well. Making logistical and funding...expect to generate data that will contribute to a publication? A grant? A patent? It is never too early to ...Resources Resources on the Addgene blog Fostering communication in remote collaborations Transferable Skills...

...unique barcodes for each TF help streamline identification. While the Zhang lab controlled their multiplicity...single TF was delivered to each cell, future applications could deliver multiple TFs to each cell, allowing...processes. For help with library transformation and amplification, refer to the Zhang lab’s previously published...published protocol and its relevant modifications on the pooled library webpage. Disease models Good disease...generate, in part because of difficulty with replicating the specific dysregulation in the disease. For...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome Library...

...rounds of replication are initiated before the cell has divided. This means that genes located near the...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes...genes closer to the origin of replication will be overrepresented compared to those further away. As a...maximal gene expression near genomic origins of replication, the stabilized promoters created by Segall-Shapiro...the control of IPTG. Because RepA is a plasmid replication initiator protein, they could alter copy number...

...red fluorescent proteins) allow simultaneous identification of more than one protein of interest. However...problematic, because the DREADD receptors need to be located on the plasma membrane to be activated by the DREADD... species (bottom panel in Fig. 1). Our results indicate that the tag protein may have an important influence...fluorescence expressed in the neuropil conceals identification of opsin-expressing cells). For other things...Rodgers L, Wilson IA, Lerner RA, Wigler M (1988) Purification of a RAS-responsive adenylyl cyclase complex...(2013) Overview of Affinity Tags for Protein Purification. Current Protocols in Protein Science 73: . ...

...members of their pathways, but where the kinases are located, their signal strength, and how long their signals...and MEK1 is active or “on.” Dronpa’s color thus indicates MEK1’s current state (Green when off, dark when...domains were inserted at sites homologous to the locations they were attached to in MEK. The photoswitchable...kinases good for? There are a number of potential applications for photoswitchable kinases.  Below are a few...signaling by using psMEK1 and an ERK kinase translocation reporter with a mRuby2 fluorescent reporter ...endogenous ERK, and its localization serves as an indicator of MEK1 activity. When MEK1 is active, it phosphorylates...

...detected with tyramide signal amplification. Tyramide signal amplification occurs when HRP converts a fluorescently...reporters (i.e. fluorescent dyes, HRP, etc. ) after purification. Antibodies are difficult for an individual ...culture media. Their His tags allow for further purification, if desired, but it's not required for their...some reporters are better suited for particular applications. Read on to learn more about each reporter or...Pros Cons Antigens HRP (P) Tyramide signal amplification of HRP Staining cultured cells and tissue sections...

...These properties allow for efficient signal amplification, making biotin conjugated antibodies good options...especially if you start to consider clinical applications). But there are a few other classes that we ...proximity ligation, immuno-PCR, and single-cell applications (Hegazy, et al., 2020; Niemeyer, et al., 2007...also conjugate antibodies directly to beads for purification purposes.  Coupling up - antibody conjugation...added to antibodies post-translationally. 4) Modifications made to antibodies, such as inclusion of non-canonical...Detecting Protein-Protein Interactions and Protein Modifications in Cells and Tissues in Situ. Curr Protoc Cell...

...days prior to the implementation date, our data indicated that, while 69% of all postdocs were expected ...sent, with the lack of transparency and poor communication of their plans (many postdocs - and in some ...to reach out to institutions with a statement indicating that we encourage them to keep raising salaries... little correlation with cost of living at the location of the institution. Continuing efforts to gather...Director of Future of Research. He is interested in advocating for and with junior scientists, to help them ... Bring Increased Postdoc Wages Read about the Advocating for Science Symposium and Workshop Learn about...

...digital notebook and make sure you note the freezer location of every plasmid in your collection. Use one of...access and use the materials they need, and avoid duplicating your effort. In addition, if you should choose... important step in ensuring quality, as it will catch any errors made during the cloning, sample handling... for more tips on sequence analysis, plasmid verification, and using NCBI BLAST. Tips for high-throughput...result against the expected sequence for quick verification, which can be done through the align tab of ... plasmid verification...

...As evidenced by all the CRISPR publications, press, and plasmids out there, it’s obvious that CRISPR ...nickase variant, lowering Cas9 expression, and truncating gRNA sequences used for targeting; however, these...on-target and off-target activity at multiple genomic locations in HEK cells. Further testing of two of the mutants...PubMed PMID: 26628643. Find plasmids from this publication at Addgene. 2. Nishimasu, Hiroshi, et al. "Crystal...PubMed PMID: 26735016. Find plasmids from this publication at Addgene. 5. Chen, JS, et al. "Enhanced proofreading...PubMed PMID: 28931002. Find plasmids from this publication at Addgene. Resources on the Addgene Blog Read...

...repurpose popular GFP reporter lines for more sophisticated experimental manipulations, saving the time ..., 98% of TdTomato expressing cells were GFP+, indicating a robust, but specific system. T-DDOGs also successfully...systems are easily adaptable to neuroscience applications, including optogenetics, and they should make...Could Cas9-DOG be developed soon to allow genome modification only in a small subset of cells? Other proteins... PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “...PubMed PMID: 26258682. Find plasmids from this publication at Addgene. 3. Kirchhofer, A., et al. “Modulation...

...choose well. What are their qualifications? Do they have certifications? How can they demonstrate their...  Check out Joanne's Reddit AMA I am a strong advocate for peer mentoring, but volunteer mentors are ...help you. A good coach usually will be a clear communicator and make direct suggestions for improvement ...coaching is nice if you can get it.  Because I am located in Cambridge, MA (and because we have a lot of ...she is especially excellent in coaching for communication skills. Industry transition is also an area ...

...experiences to undergraduate students (1, 2), but educators face significant challenges in designing suitable... become very different due to a whole genome duplication and extensive rearrangements in the S. cerevisiae...overexpression vector. This plasmid contains origins of replication and selectable markers that allow students to...met deficiencies (manuscript in preparation), indicating that Met protein functions have been largely ... research into undergraduate courses and in communicating science to the public. You can find out more...). Vision and Change in Undergraduate Biology Education: A Call to Action, Washington, DC. 2. President's...

...scientific poster apart? First, a good poster should catch the audience’s eye and draw people in. That can ... sections should be as concise as possible, communicating only the most important results to your audience...have the advantage of being able to verbally communicate your work, so you don’t need every little detail...may choose to display it as rows. It can be more catchy because it is generally less common and will stand...advantage of being different and potentially more catchy, but it can be trickier to make figures fit nicely...relevant to the work you are presenting. The latter is catchy, but beware. A distracting background or clashing...

..., its dimeric nature can interfere with some applications. Several labs have been working to produce monomeric... for molecular fusion and membrane-targeting applications. Nature Methods, 1–9. https://doi.org/10.1038...library generation and screening that led to identification and characterization of AAV.CAP-Mac. Image ...rodents and broad tropism in primates. Nature Communications, 14(1), 3345. https://doi.org/10.1038/s41467...proven itself a good option for essentially all applications. Addgene’s plasmid collection includes over ...clone (M2) has been widely used in a variety of applications. Find anti-DYKDDDDK recombinant antibodies here...

...the best, or even which one will work for your application? Like many scientists, I have spent hours sifting...produced from a new set of animals), and wrong application (different experiments/conditions can change ...differently over time), compatible with the desired applications (such as immunohistochemistry (IHC) or western...colleagues! Dr. Wu and his lab members (like me!) are dedicated to creating a public and accessible database for...expectation and comparison on the antibody signal’s location. If a distinct imaged pattern does NOT align with...literature and is unrelated to the target protein, we categorize the antibody as having non-specific binding in...

... biological reagents have a ton of data on the catalog page. It’s not always super easy to navigate the...originally described in a peer-reviewed scientific publication. We love science, but we don’t have the resources...questions about an Abcam antibody. Have your catalog number ready. I've attempted to answer several ...Addgene support, make sure to mention the plasmid catalog number, or at least the plasmid name in your initial...be fairly fast, getting the full answer to a complicated question may take some time. If the customer ...message Answering customer service questions is a delicate balance of knowledge, patience, and diplomacy,...

...chemical replication of RNA in the absence of evolved enzymatic machinery (i.e. RNA replication with no...Guangzhou and the Science City The conference was located in the modern, relatively new “Science City” area...and technology companies and start-ups choose to locate there. I made time to go into the city of Guangzhou...inheritance that might be less error prone than DNA replication. He too mentioned how “surprising” small RNAs...Steitz quipped that RNA in the ribosome is in a complicated conformation “not like DNA which has a boring...

...so most labs opt to purchase adenoviruses from dedicated adenoviral production labs or companies. Production...something you may have to optimize for your specific application. With mammalian systems, it is standard to do...more cells, while AAV can be titrated following purification using qPCR or digital droplet PCR. Insect cells...to collect your viral supernatant. Baculovirus replicates inside insect cells, and its lifecycle ends in...titer the virus, or go straight into your viral application, like protein production. Titration of baculovirus...Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation (AAV); centrifugation (lenti...

...antiviral defense). Cascade is brought to the target location using a gRNA and can then recruit Cas3, which ... type I, the most abundant type in the Class 1 category. These type Is can then be further broken into...of DNA much like a fisherman would reel in a big catch. It was originally suspected that Cas3 moved a couple...Mashimo, T. (2022). Genome editing technology and applications with the type I CRISPR system. Gene and Genome...unidirectional genome editing in human cells. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-...Koonin, E. V. (2015). An updated evolutionary classification of CRISPR–Cas systems. Nature Reviews Microbiology...