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... colors you like. Controls and validation Can’t forget the controls! The necessary control experiments...immunolabeling of brain. eLife, 8, e43322. https://doi.org/10.7554/eLife.43322 Brewer, M., McDonough, L., ...Fresh Frozen Tissue-V2 v2. protocols.io. https://doi.org/10.17504/protocols.io.bs68nhhw Im, K., Mareninov...1897, pp. 299–311). Springer New York. https://doi.org/10.1007/978-1-4939-8935-5_26 Jambor, H., Antonietti...publications. PLOS Biology, 19(3), e3001161. https://doi.org/10.1371/journal.pbio.3001161 McLaughlin, K. (2019...Immunohistochemistry. Materials and Methods, 9. https://doi.org/10.13070/mm.en.9.2846 Radtke, A. J., Kandov, E....Academy of Sciences, 117(52), 33455–33465. https://doi.org/10.1073/pnas.2018488117 Additional Resources on...

... with the Harvard GSAS Science Policy Group to organize a Minisymposium on Reproducibility. The minisymposium... additional resources and information about organizations promoting reproducibility in this booklet (which...within a published manuscript. Luckily, many organizations have been working to provide tools that make...expand to other protein technologies later. Organizations such as BenchSci and FPbase are working hard... they’ve used for data analysis while other organizations like Dryad make it easier for researchers to...are blogs - both hosted by individuals and by organizations like Addgene (a good example is our series of...editing technology NgAgo). In addition, there are organizations like ReFigure which aims to consolidate published...

...easily deliver their “cargo” to target cells and organisms, and when that cargo is designed to make you sick...biological tools In nature, viruses infect cells and organisms (which we refer to as hosts) via viral particles...and fuse with. In nature, this determines which organisms and cells they infect. In the lab, this means ...can also be directly injected into hard-to-reach organs, such as the brain. Secondly, some viruses have... makes them ideal for applications with whole organisms. Finally, viral delivery is typically straightforward...strong immune response when introduced into an organism. This feature makes them an attractive tool for...Lentiviral Transduction Additional Resources on Addgene.org Browse Our Viral Vector Pages Learn About Viral...

... cytometry. Trends Immunol 33:323–332. https://doi.org/10.1016/j.it.2012.02.010 Dennler P, Fischer E, ...Defined Reagents. Antibodies 4:197–224. https://doi.org/10.3390/antib4030197  Hegazy M, Cohen-Barak E, ...in Situ. Curr Protoc Cell Biol 89:e115. https://doi.org/10.1002/cpcb.115  Mueller BM, WrasidloO WA, Reisfeld...Monoclonal Antibodies. Hybridoma 7:453–456. https://doi.org/10.1089/hyb.1988.7.453  Niemeyer CM, Adler M, Wacker...quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/nprot.2007.267 Stoeckius M, Hafemeister... single cells. Nat Methods 14:865–868. https://doi.org/10.1038/nmeth.4380  Additional resources on the...BlottingPlasmids 101: Biotinylation Resources on Addgene.org Unconjugated antibodies at AddgeneAntibody plasmids...

...proteins are retained in the interphase and bottom, organic layer. The top aqueous layer is extracted and RNA...layer to avoid carryover of DNA from the bottom, organic layer. Another trick is to precipitate the RNA ...Biophysica Acta (BBA) - Enzymology 341:484–488. https://doi.org/10.1016/0005-2744(74)90240-x Chirgwin JM, Przybyla...ribonuclease. Biochemistry 18:5294–5299. https://doi.org/10.1021/bi00591a005 Chomczynski P, Sacchi N (1987.... Analytical Biochemistry 162:156–159. https://doi.org/10.1016/0003-2697(87)90021-2 Chomczynski P, Sacchi... years on. Nature Protocols 1:581–585. https://doi.org/10.1038/nprot.2006.83 Additional resources on the...for purifying DNA without a kit Resources on Addgene.org Check out our molecular biology protocols Find...

...In 2017, Lenny Teytelman, CEO of protocols.io, organized a panel to discuss reproducibility issues in research... there were other academics and like-minded organizations with similar ideas who were also thinking about... an eLife ambassador, had previous experience organizing open access events and grassroots efforts. After...in developing reproducibility workshops and organizational skills. Meanwhile, Addgene was also thinking... in the year and in the fall of 2018, I helped organize a workshop for community scientists at the Global... would want to know before, during, and after organizing such a workshop. Finally, the group put a lot...IS-MPMI. If you’re interested in having a workshop organized at your upcoming conference or you’d like to join...

...McKenna and Bert W. O’Malley. Survival of all organisms depends on efficient energy maintenance - through...nervous system signaling to peripheral effector organs, which then return information such as nutritional...processes are regulated at the cellular, tissue and organ level by nuclear receptors (NRs), their coregulators... of such signals between central and peripheral organs because of their established roles as molecular...activation or repression of gene expression (see www.nursa.org for a curated list of coregulators). Coregulators...context defined by ligand, cell type, and promoter, organizing the expression of functional groups of genes ...Nuclear Receptor Signaling Atlas (NURSA) at www.nursa.org. Bert O’Malley, M.D., is Tom Thompson Professor...

...cells or tissue slices mounted to a slide, or whole organs. Because the tissue architecture is preserved, ...from the bloodstream. Then, via a blood draw or organ harvest, the labeled cells can be analyzed using...and Future Outlook. Front Immunol 8: . https://doi.org/10.3389/fimmu.2017.01589 Ayyar BV, Arora S, Ravi...The nuts and bolts. Methods 116:51–62 . https://doi.org/10.1016/j.ymeth.2017.01.009 Crivianu-Gaita V, ...Biosensors and Bioelectronics 85:32–45 . https://doi.org/10.1016/j.bios.2016.04.091 Cruse JM, Lewis RE,...Immunology Guidebook. Elsevier, pp 17–45. https://doi.org/10.1016/B978-012198382-6/50026-1 Wang R, Xiang ... Skp. Front Cell Infect Microbiol 3: . https://doi.org/10.3389/fcimb.2013.00072 Additional resources on...

... 179 (2020). https://doi.org/10.1186/s12870-020-02388-2 Resources on Addgene.org Addgene’s Modular Cloning...modular cloning? Modular cloning, or MoClo, is an organization scheme for multi-part DNA assembly. Simply put...’s because MoClo specifically refers to the organizational schema described above, and not any particular...other non-Golden Gate cloning methods. MoClo’s organization schema MoClo can be broken down into three hierarchical...many guides/parts delivered) in certain model organisms. In mammals, most CRISPR libraries are assembled...targeting constructs and delivering them to these organisms via another optimized platform. Gene Stacking ...

...Girardia sp., Dugesia dorotocephala, Phagocata morgani, and Phagocata gracillis. There are 11 different...developmental biologists have to extract RNA from their organism of interest, use the RNA to create a complementary... in  the labeling specific cell types or entire organs in planarians. For example, the gene Piwi labels...strain that is designated as DE3. For Phagocata morgani and Phagocata gracillis, the process is a little...E. coli. Thus, to get the dsRNA into Phagocata morgani and Phagocata gracillis, one has to purify the ..., feedback etc. please write to cuttingclass@stowers.org and if you make new plasmids for investigating...Models for learning about CRISPR Resources at Addgene.org Find plasmids from the Sánchez lab Find plasmids...

...screens. Nature Protocols, 17(9), 1903–1925. https://doi.org/10.1038/s41596-022-00700-y Hanlon, K. S., Kleinstiver... breaks. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-12449-2 Pupo, A., Fernández,... Molecular Therapy, 30(12), 3515–3541. https://doi.org/10.1016/j.ymthe.2022.09.015 Ramani, B., Rose, I...bioRxiv (Cold Spring Harbor Laboratory). https://doi.org/10.1101/2023.06.13.544831 Santinha, A. J., Klingler...AAV-Perturb-seq. Nature, 622(7982), 367–375. https://doi.org/10.1038/s41586-023-06570-y Wang, J., Gessler, D...Transduction and Targeted Therapy, 9(1). https://doi.org/10.1038/s41392-024-01780-w Additional resources...Using the FITS Approach Additional resources at addgene.org CRISPR pooled libraries Guide to pooled libraries...

... available in your lab or organization, and approvals from the organization overseeing your work. Good...Modeling Part 2 Five Popular Model Organisms Additional resources on Addgene.org Genome Engineering Guide CRISPR...the peritoneal cavity, or the space surrounding organs in the abdomen. They are a popular choice for administering...and to avoid potential off-target pokes to nearby organs. Restraint: scruffing Location: peritoneal cavity...Biosciences and Medicines, 08(09), 131–141. https://doi.org/10.4236/jbm.2020.89011 Shimizu, S. S. (2004). Routes...

...educating the public and organizing events. I contacted local professional organizations and nonprofits. I volunteered...Central Indiana Science Outreach, or CINSO. We organize fun science events for adults and professional...communication, editing, marketing, and website organization. My short experience with my blog helped immensely...establishments such as museums, professional organizations, and other nonprofits to gain support for CINSO...read from career guidance sources such as myidp.sciencecareers.org mentioned museums. Indianapolis is home...Browse All of Our Career Posts Resources on the Addgene.org Find Plasmids for Your Research Learn About ...

...your organization Public Speaking Time management Managing a team Conflict resolution Organization Leadership... given yourself the bathroom mirror pep talk, forgotten and then retrieved your lucky socks, persevered...than you thought! When change happens at your organization, when your boss needs someone to step up, when...likely collected some great time management and organizational skills. If you are responsible for working ...students, managing inventories in the lab, or organizing a research project, one can assume that you’ve... careers Learn about mentoring Resources on Addgene.org Check out our careers page Watch our career ...

...professional works for or with an organization to evaluate the organization’s grant readiness, identify grant...stories about a time when their passion for an organization or a program and a funding opportunity coincided...submitting it, to evaluating the capacity of an organization to receive and manage funds.  Grant professionals...targeted to the grants profession? The number of organizations, both public and private, that qualify for grants...oriented nonprofits. Like Deb, I worked for an organization in a position that required some involvement...your ideal client or job will save you and the organization(s) you serve time and resources. 3. Get a mentor...

...Anti-V5 [IPI-SV5-Pk1]. Addgene Report, https://doi.org/10.57733/addgene.l0ulu4. Hanke, T., Szawlowski,...antigen. J Gen Virol., 73 (Pt 3), 653–660. https://doi.org/10.1099/0022-1317-73-3-653. PMID 1372038. Randall.... J Gen Virol., 68 (Pt 11), 2769–2780. https://doi.org/10.1099/0022-1317-68-11-2769. PMID 2445904.   ...protein degraders. Nat Commun., 15(1), 8885. https://doi.org/10.1038/s41467-024-52871-9. PMID 39406745. Optimizing...Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126.   Gene disruption in...abscessus. J Bacteriol., 206(3), e0033523. https://doi.org/10.1128/jb.00335-23.  Bioluminescent reporting...Systems. ACS Sensors, 9(7), 3489–3495. https://doi.org/10.1021/acssensors.3c02664. ...

...without DNA cleavage. Nature 551:464–471 . https://doi.org/10.1038/nature24644 Glaser A, McColl B, Vadolas...Molecular Therapy - Nucleic Acids 5:e334 . https://doi.org/10.1038/mtna.2016.48 Katti A, Foronda M, Zimmerman... Nucleic Acids Research 48:2841–2852 . https://doi.org/10.1093/nar/gkaa124 Martin AS, Salamango DJ, Serebrenik...editosome complexes. Scientific Reports 9: . https://doi.org/10.1038/s41598-018-36739-9 St. Martin A, Salamango...cells. Nucleic Acids Research 46:e84–e84 . https://doi.org/10.1093/nar/gky332 Standage-Beier K, Tekel SJ, ... Nucleic Acids Research 47:e120–e120 . https://doi.org/10.1093/nar/gkz713 Additional resources on the ...reporters Browse our CRISPR blog posts Resources on Addgene.org Browse the CRISPR guide Find CRISPR plasmids...

...into the results, it’s important to note that in Morgens et al., guides were considered toxic if they were...non-specific toxicity than non-targeting guides First, Morgens et al. looked at the effect of safe-targeting guides...guides. The genome-wide CRISPR screen presented in Morgens et al. contains three types of guides: 1) targeting... reduction in on-target efficacy. Results from Morgens et al. show that genome-wide CRISPR screens are...testing thousands of guides for off-target activity, Morgens et al drew some generalizable conclusions about...Library are available from Addgene! References 1. Morgens, David W., et al. "Genome-scale measurement of ...Guide to All Things Lentivirus Resources on Addgene.org Check out this guide to pooled plasmid libraries...

...Watkins, Professor at George Washington University and developer of wesharescience.org. Sharing your research...professor at George Washington University and he the developer of the WeShareScience.org platform. You...study processes for diagnosing and resolving organizational performance and often have to be able to quickly...sharing video abstracts about research: www.WeShareScience.org. WeShareScience is a free platform for ...Sharing in Research Publications Resources at Addgene.org Check out our Protocol Pages for Video Protocols...

...asks for strong organizational skills. You might think, “I’d be good at that! I organized my lab data so...important part of the application. Even within an organization, there can be variance. At Addgene, our content...explain why you are interested in working for that organization or group. This is particularly important for...want to talk about! You could describe your organizational system, your rationale for designing it the...title of the position] at [name of the company/organization]. I am a [adjective that describes you] scientist...that particular job, your commitment to the organization's mission, or your deep interest in the lab's...