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...It is also worth noting that plasmids can be symmetrically incompatible, meaning that both plasmids are... from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher... higher rate than the coresident plasmid. Asymmetric plasmid loss occurs for several reasons, including...high copy incompatible plasmids that lead to asymmetrical plasmid loss of the usually large, low copy ...

...enzymes (BsaI, BbsI/BpiI, BsmBI), which recognize asymmetric DNA sequences and cleave at a short distance ...plasmid stocks. To avoid cross-contamination, we recommend retrieving plasmids from your original Addgene...fragments. To increase your ligation efficiency, we recommend you calculate the right volume of each plasmid...efficiency. If you’re cloning in high-throughput, we recommend transforming into chemically competent cells in...

...therefore differ from library to library. We strongly recommend following the protocols provided by the depositing...have a specific protocol for producing virus, I recommend that you follow that protocol. If not, our protocol...polyethyenimine transfection protocol. I also recommend searching PubMed for any publications associated...ddPCR) to quality control our viral preps, we recommend determining the functional (infectious) titer ...

...neocortex of a living mouse. Image courtesy of Axel Nimmerjahn (Salk Institute for Biological Studies) ...oligomeric state of a fusion partner. Therefore, we recommend using miRFPs and their enhanced versions named...detection of protein-protein interactions, we recommend using NIR split reporters based on miRFPs (Shcherbakova...Biol 20, 1078-1086 (2013). Shcherbakova, D.M., Cox Cammer, N., Huisman, T.M., Verkhusha, V.V. & Hodgson, ...

...data. Finally, let me take this opportunity to recommend all the science and career talks at iBiology which... careful. 3. Tell a story We are biologically programmed to listen and remember better when we hear material...directions. Don’t worry if your work hasn’t been commercialized, just talking about the possibility is interesting...sharing model works. See if you can elicit helpful comments from your audience as in “Does anyone else have...

...bacteria. It’s hard to count the number of different commercial strains of E. coli currently available – a quick...deadly to humans. The majority of all common, commercial lab strains of E. coli used today are descended... E. coli strains used in the lab Most of the commercial strains you find today are marketed for a specific...make these features possible are present in most commercial strains, especially mutations that make major...

...IF labeling of rat brain samples from the James Trimmer Lab, using recombinant antibodies with different...fluorescently conjugated secondary antibodies are commercially available. Pick whichever combination of host...questions or any favorite tips and tricks, leave a comment below. Happy imaging! References and Resources...Der List, D., Murray, K. D., Engebrecht, J., & Trimmer, J. S. (2019). A toolbox of IgG subclass-switched...

...excited to do what they do. I would also highly recommend talking to other undergraduate researchers, as...point ties into the previous one too. I would recommend applying for multiple positions—don’t put all ...Experiences for Undergraduates (REUs) and other summer research opportunities are also fantastic for broadening...friends and found incredible mentors through my own summer experience. Also look into presenting your research...

... Regenerative Medicine (CRM), Edinburgh. Most commercial antibodies do not work in pull-down assays: Epitope...because reagents could be purchased directly from commercial suppliers without investing time and labour on... gene-specific crRNAs and matched ssODNs from commercial suppliers of CRISPR reagents (turnaround time...method. We anticipate that our method will open up immense possibilities to delineate the function of large...

...settings of the equipment. Systematic evaluation of commercially available dyes with different excitation wavelengths... the photons emitted. For example, DAPI is much dimmer than Alexa Fluor 488, and thus it is more difficult...Sridharan et al., 2014). Manufacturers will typically recommend specific fluorescent donors and acceptors in their...Marketing Manager at Promega. His work focuses on commercializing their cell and protein analysis products. You...

...” by Sara Zimmerman   5)   “Beatrice the Biologist” by Katie McKissick       Sara Zimmerman, Unearthed...list? Let us know by sharing it with us in the comments.   ...

... for deposits of less than 10 plasmids but we recommend using the spreadsheet option for deposits of 10... been approved. The spreadsheet option is recommended for depositing 10 or more plasmids. (If you believe... lentiviral, retroviral, and AAV plasmids, we recommend the NEB Stable strain. For plasmids containing...containing the ccdB gene, such as Gateway vectors, we recommend the ccdB Survival strain. Figure 10: Enter the...

...Readouts Toj, a summer intern who shared their experience working at Addgene. An Addgene Summer Internship...Flow Panel, and Beyond Surface Labeling Ashlyn Lemmen, who shared her flow cytometry expertise in two...

...If you have any comments about the Hot Plasmids Podcast Segment, or have recommendations for new Hot Plasmids...Plasmids topics, let us know in the comments section below. Hot plasmids covered in this segment Hot Plasmid...

...bacterial cultures (i.e. 2-3 ml) with or without commercially available kits and DNA for transformation doesn... required sizes of the bacterial cultures and commercial kits you will adopt for the extraction. Restriction...and outcomes of experiments. For these reasons commercially available kits provide extra washing steps to...

...intellectual property rights, pending publications, and commercial use often come up. An MTA can resolve those questions...that recipient may not use the material for a commercial purpose Assurance that recipient may not use...inventions Right to publish without editorial comment or review by the Provider Limited liability ...

...genomic target, you need to make sure the NGG immediately follows your target on the genome but NOT on ...considerations only matter if the PAM sequence is immediately adjacent to the nearly-homologous target sites... you have any suggestions? A14: The Zhang lab recommends using the Epicentre QuickExtract solution to ... as the second design (in pX330/PX335), so we recommend people all use the pX330/PX335 system. Additionally...

...D. W., Kumaran, R., Rego, M., Schofield, C., … Laflamme, C. (2024). A consensus platform for antibody ...10.1038/s41596-024-01095-8 Kahn, R. A., Virk, H., Laflamme, C., Houston, D. W., Polinski, N. K., Meijers,...Errington, T. M., Turn, R. E., Bandrowski, A., Trimmer, J. S., Rego, M., Freedman, L. P., Ferrara, F.,...

...comprehensive job description for this co-op, I was immediately interested in applying. This had stood out to...listed with a much less specific overview. It was immediately clear that Addgene wanted to ensure applicants...Letter 7 Tips to Secure a STEAM Internship This Summer  ...

...periodically beat their tails left and right in a symmetrical motion to move forward. This behavior is not ...Kolmer-Agduhr cells was sufficient to trigger the symmetrical tail movement unique to spontaneous slow forward... photostimulation in order to drive the full symmetrical response.  The function of Kolmer-Agduhr cells...