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...simultaneously allowing scientists to actively comment, annotate, and edit protocols. After my introduction...them public for others to use, and add your own comments. Currently you can find protocols from NEB, various...protocols to share with others. The site allows you to comment on protocols and create lists of your favorite ...software app that I missed, let me know in the comments below and leave your thoughts on how the app works...

...to maximize the alignment length. While this recommendation is not infallible, we have found it saves analysis...sequencing result and can easily translate it, we recommend using blastp over blastx. The primary advantage...confirm a tag or fusion protein is in-frame, we recommend using blastx with the "Align two or more sequences... to confirm your plasmid sequencing results or comments on our suggestions? Share your thoughts here to...

...Polyclonal Can be labeled with multiple dyes Commercially available Not recombinant; Require animal... -mouse versions are currently available Not commercially available Small but Mighty: Advantages ...and several maleimide-conjugated dyes are also commercially available. References 1. Pleiner, T., Bates...supernatant is only $40, significantly cheaper than commercially available antibodies....

...cleavage. How does SHERLOCK work? Cas13 can be programmed with crRNA to target an ssRNA of interest, for...pathogen. Once Cas13 recognizes and binds to the programmed sequence, it can promiscuously cleave surrounding...cleavage byproducts. Thus, binding of Cas13 to the programmed region of interest would lead to both the cleavage... that a cleaved reporter could be detected on commercial lateral flow strips, similar to pregnancy tests...

...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 ...

...reprogramming while PBMCs collected from blood are reprogrammed immediately after collection [2]. PBMCs display unique... fibroblasts obtained from aged patients are reprogrammed less efficiently, no such correlation has been...An Efficient Protocol Using Serial Plating of Reprogrammed Cells by Centrifugation. Stem Cells Int, 2016...

...references organized from the beginning. I also recommend using the citation style of (Last name, Year) ...about having to hunt down sources later on.  A comment about writing: initially I felt that I needed a...get done between office hours and experiments. A comment about citing: I wasn’t sure if I should cite reviews... be revolutionary, but you could include a few comments on where you think the field is going or what ...

...highlight these recent advancements and offer commentary on their application to neuroscience research...To highlight this advantage, they selected the immediate-early gene Bdnf, which encodes brain-derived neurotrophic...early in response to neuronal activity (hence, “immediate-early”) and consequently regulates neuronal physiology...and Bdnf IV overexpression variants were other immediate-early genes associated with neuronal activity,...

...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 ...

...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...

...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...

...to people outside of academia. Here are my recommendations on some ways to get started sharing your science...others in the science blogosphere is to read and comment on their blogs. 5)   Local science museums are ... research by guest blogging here, just leave a comment if you're interested and our team will get back...

...PITChing MMEJ as an Alternative Route for Gene Editing Michohomology-mediated end joining (MMEJ), also ...favorite CRISPR tool or resource? Let us know in the comments section!...

... hepatic or intestinal cell types. MEFs were reprogrammed into iEPs by retroviral expression of transcription...a ‘dead-end' path where cells were partially reprogrammed and re-expressed fibroblast genes. Most cells...cocktail yielded a three-fold increase of fully reprogrammed cells based on single-cell gene expression patterns...

...antibodies available at launch were created by James Trimmer, PhD, of the University of California Davis School... Figure 1: Antibody against PSD-95 from the Trimmer Lab. Information about the antibody, the target...Nguyen K, van der List D, Murray KD, Engebrecht J, Trimmer JS (2019) A toolbox of IgG subclass-switched recombinant...

...the CO2 goes out. Conclusions and handling recommendations Not all O-rings function optimally at -80°C...results and the results reported elsewhere, we recommend that once you receive your AAV from Addgene, you...to thaw. However, if this is not possible, we recommend both venting the tube for at least 30 seconds ...

...onto the smear until the alcohol runs clear and immediately rinse. Flood the smear with the safranin counterstain...blot gently to dry, and then view under an oil immersion using a light microscope. Gram positive bacteria...a lab.   Anna Kamens is a science communication summer intern at Addgene and a rising third-year at Old...

...are multiple algorithms, both open-access and commercial, that are commonly used for the prediction of...common open-access tools, while EpiMatrix is a commercially available algorithm to evaluate the immunogenicity...doi.org/10.1016/B978-0-12-417144-2.00027-5van Brummelen, E. M., Ros, W., Wolbink, G., Beijnen, J. H., ...

... Roth Lab recommends verifying cre expression prior to use. The lab additionally recommends using an anti-mCherry...effect doesn’t confound your experimental data, we recommend generating a monoclonal Cas9-expression line prior...

...blot out quickly." Final design (right). Immediate and immense regret of the graphic designer not shown...microscopy and flow cytometry experts for the panel recommendations. A few discussions in the review process led...

...conjugates, blocking the conjugation reaction. Many commercial antibodies come with sodium azide already included...reaction is done, through a variety of methods or commercial kits.  However, antibodies conjugated to horseradish...use it for your ELISA and Western, I’d highly recommend sticking with what works.  To balance this out...

...economically disadvantaged students for paid summer internships. A summer teacher professional development program...with both of these resources road tested over the summer with dozens of teachers around the country. How...

...but plasmid-based SDM has lagged behind. While commercial kits are available for making small point mutations...plasmid, which could introduce mutations. Using commercially available high-fidelity polymerases such as ...as KOD and Phusion, which are recommended for plasmids up to 25 kb, most cloners should be fine, especially...

...reviewer’s comments without explaining why. If it looks like you have ignored their comments, they will.... In addition, authors and readers are able to comment on the referee reports. If the reviewers suggest...

...consensus recommendations, even when a federal mandate encourages them to do so. Immediately after the... with raises, as for years there have been recommendations that postdoc salaries be raised to $50,000 ...

...connecting. You can see what they post and occasionally comment on their posts as appropriate, possibly giving ...Keep it professional at all times Do not post or comment on anything that is not relevant to business or...inappropriate content see this article. However, do comment on other people’s news and updates. Share your ...

...reactions. After all, there is an abundance of commercially available conjugated antibodies that are ready...talking about this? Even though there are so many commercially conjugated antibodies available, that never ...antigen recognition and affinity. Hopefully, a commercially available conjugated antibody will have been...

...consider the reviewer's comments. Whether the proposal is funded or not, the comments help gauge room for ...include for the next application. Although some comments may make it clear that the reviewer misunderstood...

...optimization. Before beginning an experiment, it is recommended that you try a few different ratios of transfer...will need to be optimized prior to starting. We recommend calculating the ratio of the surface area of the... target cell line. To overcome this issue, we recommend using a range of doses of lentivirus when transducing...

...you can get one. As a starting point, I would recommend conferences like Lab Robotic Interest Group (LRIG...machine fits on a table top and can be easily programmed for a variety of lab needs. Addgene uses it to... tips and tricks, be sure to share them in the comments section.   Additional Resources on the Addgene...

...scientists, I have spent hours sifting through comments and papers to find the most reliable (and cost-effective... crisis” came to light as antibodies became commercialized, revealing the unreliability of antibodies....antibody crisis states that up to 50% of all commercially available antibodies are unreliable (Baker, ...

...trying to strip and reprobe. In this case, we recommend using mild stripping solution (see below) to minimize...Run your gel and membrane transfer as normal. Immediately after transfer, dry the membrane to maximize ...retention. Following this step, most PVDF membranes recommend re-wetting with methanol before rinsing with wash...

...takes on the order of minutes. Studies looking at immediate effects of specific neuron firing are better equipped...protein-coupled receptors (GPCRs), that have been reprogrammed to respond only to inert, nonnative ligands....Mouravlev A, Carpenter JC, Snowball A, Knauss S, von Schimmelmann M, During MJ, Lignani G, Schorge S, Young D,...

...software tools they recommend — feel free to share your favorite software in the comments below! References...2019). When possible, the PE3b/PE5b approach is recommended over the PE3/PE5 approach. Figure 7: ...