...be precise, the fixing solution is different while the permeabilization solution can be the same). Examples...can be achieved through two-protein labeling by utilizing(a) biotin-streptavidin binding and (b) antibody...then permeabilized, using two different buffer solutions. Fixing causes protein cross-linking, killing ...species reactivity. When sorting cells with a high resolution for specific downstream applications, while maintaining...

...resultant gene targeting system could be used to routinely deliver sequences of interest to cells with efficiencies...CRISPR editing CRISPR, of course, has started a revolution in gene editing. However, while CRISPR is remarkably...the AAV approach is most well suited, since it routinely generates knockin efficiencies of 1-40% with G418...PMC4752446. 5. Asuri, Prashanth, et al. "Directed evolution of adeno-associated virus for enhanced gene delivery...nucleases with adeno-associated viral vectors for therapeutic gene editing." Precision Medicine, CRISPR, and...

... Returning to an earlier analogy, no amount of shouting will make an apple fall up. As scientist and politician...GMOs), the absence of a link between vaccines and autism, and other hypotheses mount yet policies and public...proposing to defund aspects of research at National Aeronautics and Space Agency, the Environmental Protection...Transgender issues Reproductive health Immigration Pollution Healthcare Science education To make scientists...and inclusion through funding and at their own institutions, while many also seek to honor and credit women...

...have waited. Both tasks have some utility, but I lost all the utility (fun) of eating with friends by splitting... two main things: How useful the task is (its utility) How much time it will take you to complete the...should err on the side of things that have the most utility; i.e. those things that will be most useful for...and think about what they mean in terms of the utility of your tasks. In my cell splitting anecdote, I...

...these issues, the Ting lab utilized yeast display-based directed evolution to generate two mutants, the...the Brian Burke Lab (Roux et al., 2012). BioID utilizes the E. coli biotin ligase, BirA*, with a site ...Thus they conducted multiple rounds of directed evolution until they identified a mutant with significantly...PMC4831873. Lam, Stephanie S., et al. "Directed evolution of APEX2 for electron microscopy and proximity...

...exist for this, the most common of which is the utilization of phosphomimetic residues. The phosphorylated...researchers with a valuable tool for understanding the contribution of phosphorylation to enzyme function. The development...tRNA that recognizes a specific ‘open’ codon and utilizes this triplet to incorporate phosphoserine A Sep-tRNA...events on its activation loop, but can be made constitutively active through phosphomimetic mutation of these...phosphoserine) to control for effects of the amino acid substitution. Alternatively, you can express the completely...

...clear on his science, and found a solution. However extreme a solution may seem, you should consider it...greater than $5,000 but this can depend on the institution/funding source. Major equipment includes large...your research training to help you find other solutions to reach your goals. For example, a junior investigator... point of a negotiation is to create a win-win solution for the success of your science. If the resources...

...Promega’s Advanced Technologies group as well as evolution of the coelenterzine substrate to the more stable...making it non-optimal for in vivo imaging. The solution is to couple Nluc with a red-shifted fluor. Here...other cellular proteins. BRET-based biosensors utilizing NanoLuc® luciferase Many intracellular sensors... from Addgene and we're excited to see how you utilize them in your group! Many thanks to our guest blogger...Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem ...

...of all our depositors here at Addgene — your contributions make the repository what it is. So we’re thrilled...strikingly similar to the design model at atomic resolution” (Kuhlman and Dantas et al., 2003). That made...folding and selective pressures during protein evolution. The Baker Lab has deposited multiple Top7 plasmids... at Addgene, along with other tools like high resolution Ras sensors and crosslinked peptides proposed...

... the University of Manitoba. Our lab has been utilizing chemogenetics (Designer Receptors Exclusively ...fact that AAVs themselves are not pathogenic, precautions should still be taken when using them. There ...resistant to alcohol-based disinfectants. A 10% bleach solution should be used to clean AAVs from workspaces. ...being made to make them even more useful for therapeutic purposes. The University of Pennsylvania and ...Cas9 but have the potential to deliver other therapeutic molecules such as microRNAs (miRNAs), short hairpin...

...in pull-down assays: Epitope tagging provides a solution Proteins - the workhorses of the cell – never ...expensive, and not to mention frustrating. One elegant solution to sidestep the antibody problem is to tag the...tagging. Using our 96-format pipeline, we can routinely achieve 5-30% tag knock-in efficiency across different...experiments using the csRNP method is as simple as doing routine PCR assays. Here is a simple to-do-list to follow...should be pursued, however, clonal lines must be cautiously analyzed for any damage (indels) to the promoter...

...promoter regulating the gene of interest with a constitutive, robust M24 promoter. Originally isolated from...from the Mirabilis mosaic virus, the constitutively active M24 promoter has been modified with duplicated...large gene transfer. The M24 promoter provides constitutive, high-level gene expression and is functional...antibiotic resistance gene is controlled by a weaker constitutive Nos promoter, and it can be used to select both...

...Designer Drugs): Chemogenetic Tools with Therapeutic Utility. Annual Review of Pharmacology and Toxicology...Optogenetics offers a high degree of temporal resolution as receptors can be turned on and off simply ...sustained activation period is beneficial for use in therapeutic or behavioral applications where drawn out effects...

... shy away from looking beyond its confines. The beauty of serendipity often emerges at the intersections... key to unlocking new perspectives or even revolutionizing your research. Why serendipity matters Serendipity...lead to unexpected discoveries and innovative solutions. Allowing for serendipity in your lab research...benefits by encouraging activities beyond the lab routine. It encourages you to think beyond the confines...PhDs in their careers with actionable steps and execution plans.    More resources on the Addgene blog ...

... a team to get a lot of stuff done.  Factors contributing to job satisfaction Being a good manager is ...organization and feel the importance of their contributions. Flexible work schedule Flexibility to manage...personal growth do not exist or if they feel underutilized in their present positions By removing the...will address more specific tactics managers can utilize to be in touch with the needs of the team and to...

...structural modeling and structure-guided direct evolution, Nathan Shaner and colleagues from The Scintillon...Karolinska Institute developed mNeonGreen via 21 substitutions to LanYFP and the addition of the enhanced GFP... to FRET and stochastic single-molecule super resolution imaging. mNeonGreen also has little sequence ...Addgene! References Ai, Hui-wang, et al. "Directed evolution of a monomeric, bright and photostable version...

...to complete your story: an introduction, and a resolution.   Fig. 2: The structure of a story - ... left out, which can be a big challenge!  The resolution Throughout your conflict, you’re encouraging ...audience - via the scientific method - to the resolution, or the ending of the story.  At this point you...complex story, and then build on that in your resolution to really dig into the significance of the results...

...communication graduates are often well-equipped. Many institutions are sincerely interested in connecting with ...of how to appeal to a given audience and how to utilize a wide array of media in doing so—can prove invaluable...risk perception, scientific uncertainty and precautionary regulation was all necessary to inform the experimental...unsettled science. Understanding just how the public utilizes insights gleaned from scientific research is, ...

...deftly coordinated by the eukaryotic cell for the execution of many complex cellular behaviors, such as differentiation...engineering realm of S. cerevisiae and E. coli. The utility of simultaneous and multidirectional multiplex ...off switch The multiple scRNAs all individually utilize the same dCas9 protein. By using an inducible promoter...Western Ontario studying plant-based recombinant pharmaceuticals. Follow him on Twitter @AdamChinFatt     References...

...treatment options, as well as non-pharmacological solutions to modify the course of this condition are urgently...glowing bacteria and a fish robot to detect water pollution Dr. Julio Ernesto Valdivia Silva, Universidad ...related to space medicine. Now he aims on tracking pollution in rivers by employing synthetic biology resources...Biology Resources Learn about Global Plasmid Distribution ...