... added on a regular basis, so the ecosystem of related tools is always growing. Beyond the ability to ...formatting. In this way data is moved from a static and isolated vehicle and becomes part of the dynamic and connected...understand and/or attempt to reproduce research relating to data deposited into a repository access to ...can be viewed – and conceivably additional work relating to reproducing the original research can be documented...

...binding specificity: SpCas9 variant Mutations (relative to SpCas9) PAM sequence D1135E variant ...bacterial species Many more Cas9 orthologs have been isolated, and our understanding of their PAMs is shown ...same species. Expanding the CRISPR toolbox The isolation of novel CRISPR proteins has and will continue... an intrinsically high-fidelity genome-editing platform. Genome Biol 19: . https://doi.org/10.1186/s13059...

...it helps you later on to remember where you read particular studies or experiments. Later, you can easily...have to worry about having to hunt down sources later on.  A comment about writing: initially I felt that... all the proteins in the class that showed the relative sizes and the conserved domains. This information...

...Adenoid Cystic Carcinoma, Up-Regulation of pVHL along with Down-Regulation of HIF-1α by NDRG2 Expression...a cell line even decades after it is initially isolated. Regrettably this is not usually the case if you...official biorepositories or have been directly isolated from patients. In these cases, a test known as...

...bi-directionality. For predominately anterograde tracing, biotinylated dextran amines (Veenman et al., 1992) or fluorescent...al., 2007; plasmids at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have...overview see Addgene’s AAV guide. AAV particles are relatively small (20 nm) in size which makes them capable...Transport: Cargo-Specific Mechanisms of Motility and Regulation. Neuron 84(2), 292–309. PubMed PMID: 25374356...Veenman, C.L., Reiner, A., and Honig, M.G. (1992). Biotinylated dextran as an anterograde tracer for single-...

...requires time and knowledge. In a recent article, we translated our knowledge of electronic lab notebooks into...ELN should allow you to collaborate within the platform and avoid countless emails. Important collaborative...permissions to different lab members to avoid any issues later on. Do you need external teams or partners to access...lab (user, date, time, action) are required for regulatory purposes in many labs. Besides this being of ...

... design and engineering of fusion proteins is relatively straightforward. However, generating a fusion...sites or motifs that are necessary for post-translational modifications. Despite these potential issues...fluorescent protein. Thus, scientist began using relatively long linkers between the protein of interest ...C-terminal residues of the fluorescent protein are relatively close together (figure 1). Where should the fluorescent...

...color, allowing for the fate mapping of cell populations over time. The ability to track single cell dynamics... possible from Brainbow. Tracing clonal cell populations using the Confetti mouse For mammalian systems...not present at the same frequencies. The small population size of CFP++ cells permitted researchers to ... and Lgr5 identify two functionally distinct populations." Proceedings of the National Academy of Sciences...

...fractionation kit or a lysing protocol developed for isolating proteins from a specific compartment. If your ...blockers, you can run a comparison of different formulations instead of concentrations. Most labs use protein-based... For nitrocellose membranes, you can even give gelatin a shot.  Remember that protein-based blockers degrade...in an assay, meaning that your normalization calculations will not be correct, and you’ll need to adjust... Splice variants Check literature Posttranslational modifications Check literature Highly...

...harassment Isolation – including preventing access to opportunities, physical or social isolation, withholding... questions in the sections below. Mentoring relationships can help identify your priorities and needs,...regretted it when the same thing happened to her a year later. I hear too many science trainees get warnings and...

...certain region or calculate a colocalization factor to assess the spatial relationship between the targets...need more signal, another strategy is to use a biotinylated secondary antibody then label that with fluorescently-conjugated... other antibodies are disrupting the binding of later targets, the sample is being damaged by all the ...

... day or a week planning an experiment, even a relatively simple experiment. However, by taking this time...surprising findings when studying a seemingly unrelated problem. However, you have finite time and resources...background slides. If you don't have much new data, speculate on what you hope to accomplish and how your research...satisfying. Finally, many PFs are in committed relationships and/or have started a family. Those people are...

.... Speed: AAV transduction experiments are relatively quick. We have found that expression occurs quickly...AAV selection, there are three important factors related to location. Location (1): What tissue? Location...tools for neural tracing in very specific cell populations. Find Ready-to-Use Retrograde AAV Preps at Addgene...and Mark A. Kay. "Future of rAAV gene therapy: Platform for RNAi, Gene Editing and Beyond." Human Gene...

...sequence but not the amino acid sequence of the translated protein.  These changes can be 5’ or 3’ of the...strains, expression of exo, beta and gam is tightly regulated by the endogenous phage promoter pL and repressor...for a mobile recombineering system, but tight regulation of expression is required for a successful experiment...endogenous red promoter pL and the cI857 repressor to regulate expression. An antibiotic can be used to select...

...hope that these perspectives, the optimism they encapsulate, and the solutions they promote help motivate...infertility or to argue against unnecessary clinic regulations, evidence-based research is vital in combating...Greenbaum Science and evidence-based policy and legislation hit home hard in Michigan, where programs like...worried that, because we were speaking out against legislators and because the march had been so well-publicized...

...The machine is useful for distinguishing cell populations that have been tagged with different fluorescent...The optimization of GFP and its derivatives by the late Roger Tsien changed everything. For the first time...between 1,000 nm and 1,200 nm, where there is relatively little tissue absorption, weak tissue scattering..."Monitoring of blood vessels and tissues by a population of monocytes with patrolling behavior." Science...

...structural changes in the ribosome during protein translation (Ermolenko et al., 2007). All that is required... UAG) engineered into a desired site. During translation, the novel tRNA/unnatural amino acid finds the...which 50% FRET occurs. b Range corresponds to calculated D/A distances for observed FRET efficiencies ...applications." Current Chemical Genomics and Translational Medicine 3.1 (2009). PubMed PMID: 20161833. ...

...celebrate the 12 Days of CRISPR with some of the latest and greatest in CRISPR tools and resources! Ready...gave to me: Enhanced prime editing systems by manipulating cellular determinants of editing outcomes🎵 ...variants for targeted RNA degradation with minimal collateral effects🎵 On the 1st day of CRISPR, Addgene gave...

...LinkedIn. We are also exploring other social media platforms with active scientific communities and welcome...functionality and features that previously made the platform a mainstay of conversation in the scientific community...considering a gradual move off X as a response to major platform changes, such as limiting access and increased...

...Abhi Aggarwal gets the reader up-to-date on the latest version of the iGluSnFR glutamate sensorRUBY-Red...CRISPR guide library design.  dCas9-directed DNA Demethylation: Guest blogger Daniel M. Sapozhnikov tells us...modifications!) as a CRISPR/dCas9-based tool for DNA demethylation. AAV Q&A with Tim Miles: While not technically...