...would be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme variant tag that covalently...several advantages to using these self-labeling proteins over traditional fluorescent protein tags, such...endogenous target, and thus no genetically encoded tag is required. These dyes can also be attached to an...ligands have more flexibility due to the affinity tag they typically bind to. Stability of fluorescence...experimental labeling. Size is also a factor. Bulkier tags, such as GFP, may interfere with protein expression...fluorescent experiments, such as using fluorescently tagged ligands or intermolecular FRET for ligand-receptor...should possess high specificity to ensure accurate tagging. In Fluorescence Resonance Energy Transfer (FRET...

...problems resulting from past practices and mores.  His list is dire:  The current system is unsustainable... be in academia to make change.  He talked about his own unique way of funding basic research at NEB and... of the Novartis industry postdoctoral program.  His strongest comments were about the astounding lack...alternate training needs to be provided to take advantage of all the possible science careers. She advocated...

...fusion protein consisting of sfGFP fused to a His-tagged ABC transporter known to bind to the HMO of interest...precisely spatio-temporally controlled as a key advantage over other inducible enzyme technologies. She ...

... Dr. Deisseroth has also generously made many of his optogenetic constructs available through the vector... at the University of North Carolina) that offer his constructs. Importantly, control viruses (not expressing...guest blogger Derek Simon! Derek P. Simon received his PhD in Cellular and Molecular Biology from the University...Mary Jeanne Kreek at the Rockefeller University.  His interests include the role of learning and memory...is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow him on Twitter...and for good reason. For the first time in the history of neuroscience research we have a technology that...promoter of the cre recombinase while still taking advantage of the robust expression via the promoter in the...

...affinity for calcium". Please describe any tags or fusions (e.g. His, FLAG, EGFP, etc) on your insert. Use ... have the option to indicate separate promoters, tags, or cloning information for each insert.  Encodes...insert into it. Empty backbones often contain epitope tags or fusion proteins that are appended to new inserts... indicate the gene cloned into your plasmid, any tags or fusion proteins contained in the plasmid, any...Use the drop-down menu to indicate whether the tag is on the N or C terminus and whether it was present ...

...embraces open science, self-publishing directly on his website (and often posting updates on social media... vision of passive impact,” he says, noting that his team regularly shares troubleshooting tips and works...works with other labs to help them adopt the tools his group develops. And though it took a lot of work ...  Figure 1: Fluorescence intensity of EGFP-FlavinTag in E. coli changes in response to a ~25 mT magnet...effect (many do). Fusing EGFP to the self-labeling FlavinTag made a reliable one-component magnetoresponsive... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants...significant gains! After a few more rounds of mutagenesis, they eventually found a version of AsLOV2 with...

...pAAV_hsyn_NES-his-CaMPARI2-WPRE-SV40 Biosensor Eric Schreiter AV-1-PV4704 101064-AAV1 pAAV_hsyn_NES-his-CaMPARI2... Schreiter AV-1-PV4705 101061-AAV1 pAAV_hsyn_NES-his-CaMPARI2-F391W-WPRE-SV40 Biosensor Eric Schreiter...James M. Wilson AV-9-PV2453 45185-AAV9 AAV-EF1a-BbTagBY Control Joshua Sanes AV-9-PV2454 45186-AAV9 AAV-EF1a-BbChT...

...guest blogger Derek Simon! Derek P. Simon received his PhD in Cellular and Molecular Biology from the University...Mary Jeanne Kreek at the Rockefeller University.  His interests include the role of learning and memory...is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow him on Twitter...several groups [4, 5, 6]. These have the obvious advantage of not requiring patch cable connection to the...

...presented with a physical model of his large ribosomal subunit, he kept sliding his palm across the surface of... about Jacob’s initial reaction to this model on his blog. Or, for a description of how effective this... of proteins using 3D printing technology. The tagline for our company is “…where molecules become real...Addgene.org Visit Our CRISPR Guide Page CRISPR/Cas9 History Browse All CRISPR Plasmids and Resources ...

... Cloning of basic parts (promoters, UTRs, genes, tags, etc.) into MoClo-compatible vectors. Level 1: Assembly...this cloning technique! Many thanks to Rob Hurt for his help on this piece. Rob is postdoc in Peter Schultz...

...guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology and Immunology from...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz...Research Campus of the Howard Hughes Medical Institute. His interests include the quality control of secretory...Growth of new livers in culture could resolve this shortage." What is the knowledge gap you plan to address...

...epitope-targeting antibody that is conjugated to a biotin tag but not a fluorophore. In a separate step, cells ...interest. Indirect labeling is achieved through protein-tag or protein-protein interactions, like biotin-streptavidin... lab of Arlene Sharpe at Harvard Medical School. His projects include investigating negative regulators...target marker, allowing for its detection. The advantage of this system is that streptavidin conjugates...the primary antibody (Figure 1b). As above, the advantage of this system is increased flexibility for panel...of such targets include transcription factors, histones, and DNA repair enzymes. Note that when using ...stepwise decrease in fluorescence intensity in a (histogram) flow plot (Figure 4b). Through this method, differences...

...promoter. The cloning procedure adds both V5 and His6 epitope tags to the C-termini of Met proteins, which can...) from the two yeast species into the pYES2.1/V5-His yeast overexpression vector. This plasmid contains...

...out yet?) your research. Derek P. Simon received his PhD in Cellular and Molecular Biology from the University...Mary Jeanne Kreek at the Rockefeller University.  His interests include the role of learning and memory...is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow him on Twitter...reinforcement are ICSS and place preference.   Historically, ICSS was done by implanting an electrode and...technology is improving and may one day reach the sophistication of mouse transgenics. Everything that has been...the ferrule implants and patch cables. One huge advantage of using rats is their superiority as behavioral...

...jump into industry.” In his article On Leaving Academe, Terran Lane is asked by his colleagues and friends...learn about the diverse options and how to take advantage of them early is one way to smooth your path to...

...University studying mammalian immunology, and in his free time, he is pursuing a plant engineering project... cell culture and transgenesis he wants to share his knowledge with the community by teaching a DIY plant... entrepreneurs using our space to perform early-stage experiments. For example, Dave Grawoig is using ...

... for the CZ Biohub project by Manuel Leonetti to tag all 22,000 protein coding genes in the human genome... at the Biohub in San Francisco. Before starting his career in software, he completed a MSc in Psychology... tools which were optimized for HDR, except for TagIn. HDR requires the additional step of mutating guide...cut-to-insert distance for HDR. (The notable exception is TagIn––see their "summarisation score".) For CrispyCrunch...HDR. Optimal mutation in-depth One the most sophisticated features of CrispyCrunch is how it mutates guide...

...-28b-Cas9-His 6xHis (C terminal); NLS (C terminal) 62933 pET-Cas9-NLS-6xHis 6xHis (C terminal...Plasmid ID Plasmid Name Tags 69090 pMJ915 MBP (N terminal); 6xHis (N terminal); 2xNLS (C terminal... be selected, since it cleaves at the stop codon TAG. Design your repair template with 55-100 base...; NLS (C terminal) 62934 pET-NLS-Cas9-6xHis 6xHis (C terminal); NLS (N terminal)   Easi-CRISPR...

...’re attending SfN2019, stop by booth #2163 to say hi and learn more about Addgene’s viral service. Additional...topic as shown in the graphic below. Follow the hashtag #AddgeneGoesViral to find information on the basics...

...expression of the HIS3 gene, which is required for histidine manufacture and therefore growth on his-deficient...Post About Yeast Vectors Read our Blog Post About Tagging Yeast Genes ...activator (1a) or a repressor (1b). Scientists took advantage of these discoveries to create a new system for...that repress instead of activate transcription. The HIS3 reporter system (e.g. the Gateway-compatible bait... is an example of the former. In this system, histidine-auxotrophic host strains are used to select for...subset based on a cellular compartment or growth stage). A prey library can either be applied to the bait...