We narrowed to 838 results for: tat
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TypeBlog Post... to master ddPCR. Finally, it’s a good idea to state why you’re applying to the specific company or lab...rationale for designing it the way you did, and state that it would help you manage a large number of ...yourself and your background. It’s a great place to state why you’re applying to the job, company, and/or ... talked about it yet, it's also a good place to state why you are interested in working for this lab and...informal poll of Addgene hiring managers. All of them stated that a good cover letter, one that explained how...
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AAV Titers: Where Do They Come From And What Do They Mean?
TypeBlog Post...methods: Quantitative PCR (qPCR) Digital droplet PCR (ddPCR) Historically, a quantitative DNA hybridization...to report that over 95% of viral capsids in representative viral vector preparations are genome-containing...1. Fagone, Paolo, et al. "Systemic errors in quantitative polymerase chain reaction titration of self-...High-throughput droplet digital PCR system for absolute quantitation of DNA copy number." Analytical chemistry 83.22... -
Supporting Reproducibility with a Connected ELN
TypeBlog Post...characteristics and capabilities an ELN needs to facilitate true reproducibility. The following are critical...making it easy to get data out of the ELN; and (2) facilitate inter-operability with other research tools and...describe how each of these kinds of connectivity facilitates reproducibility. Connectivity to other data sources...original formatting. In this way data is moved from a static and isolated vehicle and becomes part of the dynamic...Framework, Data Dryad, and Figshare. Any ELN that facilitates reproducibility has to make it easy to export... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog Post...transfer’. Resistance can also arise from random mutations in the genetic sequence that happen to be beneficial... if some didn’t develop, either through random mutation or horizontal gene transfer, means to defend themselves... the only method of determining drug resistance status, it is a very common, well-trusted method that ...smarter, not harder. We can utilize tactics like rotating antibiotic treatments, so that exposure to each...effective drug targets (ones that are less likely to mutate or change in order for the bacterium to survive... -
The Challenges of Cell Culture
TypeBlog Post...life science literature is a litany of false interpretation in cell culture for this reason. Unreproducible...Required for Acquisition of Anoikis Resistance and Metastatic Potential in Adenoid Cystic Carcinoma, Up-Regulation..., papers are retracted and there is a loss of reputation that can impede collaborations, especially with...between cells lines, as this can be dependent on the mutation rate of the cell line, but it is broadly recommended...2010. 2. Gold, Michael. "A conspiracy of cells." State University of New York Press, Albany (1986). 3. ... -
Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs
TypeBlog Post...non-fluorescent, and misfolded FP-fusions on quantitative imaging experiments and cell physiology in general...population is not readily apparent and can confound quantitative imaging experiments or even negatively impact...misfolded dark FP fusions including erroneous quantitation of fusion protein levels. It remains unclear...fluorescent proteins and were re-engineered by mutating cysteine residues and N-glycosylation consensus...proteins. Our recent publication illustrated the quantitative increase in fluorescent signal achieved when... -
Tips for Using FRET in Your Experiments
TypeBlog Post...distance between the two fluorophores and their orientation to one another can affect FRET. When it comes...between the two fluorophores and their spatial orientation will be fixed. The quantum yield of the donor...between the two fluorophores is linked to their excitation spectrum overlap. If you choose a pair too close...increased. This method is straightforward and quantitative, but it is destructive and cannot be used for...method is also less sensitive to direct acceptor excitation artifacts and it is possible to use a non-fluorescent... -
Five Popular Model Organisms, Part 2
TypeBlog Post...conditions. As a facultative aerobe, it can thrive with or without oxygen. Additionally, its status as a Biosafety...used as a model organism for over 60 years with a status as the most studied prokaryotic organism. With ... biofilms, and much more. Because of it’s BSL-1 status, E. coli can be used as a model to relate these...plant species and metabolic properties affecting adaptations. Arabidopsis thaliana flowers. Image from... -
Five Popular Model Organisms
TypeBlog Post...different mutations found in leukemia, the Lowe lab was able to identify a set of mutations that made ...Lowe’s lab utilized mice to study how different mutations in leukemia impact different treatment regimens...by a specific gene. For example, several genes mutated in human cancers have a homolog involved in yeast... yeast: A model organism for studying somatic mutations and cancer." Nature Education 1.1 (2008): 183.... -
Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection
TypeBlog Post...genetically encoded fluorescent probes. One of the limitations is the size of GFP: ~240 amino acids or about...supplementary figure C). However, these assays are qualitative. The Organized Smooth Endoplasmic Reticulum (...the appearance of OSER whorls. This assay is quantitative and is currently the standard for assessing ...straight fusions, without any linker. When both orientations are available, a direct comparison of these ...PMCID: PMC3324619. Cranfill, Paula J., et al. "Quantitative assessment of fluorescent proteins." Nature ... -
New Optogenetic Tools for Cytoskeleton and Membrane Control
TypeBlog Post...in cells, they have various disadvantages and limitations – for example, microtubule-destabilizing drugs...damage other cellular structures, and genetic mutations are not easily reversible. Late last year, the... et al., 2022), that overcomes many of those limitations (Figure 1). Figure 1: Opto-katanin induces fast...reversible microtubule severing and has very low dark-state activity and toxicity. All of that means this light-powered...moderately-enhanced 10x and 30x SuperPLDs avoid dark-state activity and cytotoxicity better than the 50x and... -
Using AAV for Neuronal Tracing
TypeBlog Post...also use different cytoskeletal machineries to facilitate transport: retrograde transport relies on dynein...2014). It is currently unclear which proteins facilitate transport of neuronal tracing compounds and this...-Gold (Schmued and Fallon, 1986) are used. To facilitate cellular uptake of these dyes in vivo, microinjection...a negative impact on cellular health. Another limitation of traditional tracing compounds is that their...rAAV2-retro’s altered amino acid sequence might facilitate interaction with uptake or intracellular transport...mechanisms of intracellular transport of AAV could facilitate the development of more powerful AAV serotypes... -
A History of Genome Engineering in Popular Culture
TypeBlog Post...genome and are inefficient at introducing point mutations. In 2016, the researchers at the Broad Institute... envisioned a new strategy to introduce point mutations in the genome. They fused a catalytically impaired...a tRNA deaminase enzyme, TadA) to induce a A→G mutation at the target genomic loci. Together, CBE and ...the prime editor can install all possible point mutations, delete short stretches of DNA and even insert...doi.org/10.1038/nprot.2013.143 Seligman LM (2002) Mutations altering the cleavage specificity of a homing ... -
With an Eye Towards the Future, We Look Back at the March for Science
TypeBlog Post...need representatives and policymakers who not only believe in the separation of church and state, but ...Institute, most women of reproductive age live in a state with at least two abortion restrictions that conflict...are that we, as a society, need to: 1) use more citations for out data so our sources are readily available...the Vancouver SciCATs. Follow her on twitter @Point_Mutation, and @SciCATSyvr. Additional Resources on ... -
Inside Addgene's 10 Year Anniversary Gala
TypeBlog Post... aside for the event. As guests arrived, some gravitated towards the sushi and drinks, while others headed...hour, we feasted on sliders, hot dogs and a mashed potato bar (complete with all the fixings), as well as...ending the celebration. The party later moved to State Park, where I had a great (maybe too great!) time...last 10 years @Addgene Check out our impressive stats from 10 Years of Plasmid Sharing Or read more about... -
Selecting Your Plasmid Purification Kit
TypeBlog Post...are the same. Your downstream application will dictate how much DNA you need to recover and the level ...left-over debris that carried over from the precipitation stage, and then eluting the DNA from the substrate...lysis, but you will still face column capacity limitations since the columns can be “clogged” by excess ...vary from kit to kit, check the manufacturer’s documentation **Varies by plasmid copy number Pro tip! If...lots, then reviewing their quality assurance documentation can help you select a kit. So...which kit should... -
Fostering communication in remote collaborations
TypeBlog Post...final version of the document. Set and manage expectations When the researchers involved in a project are... crucial that everyone meets early on to set expectations for the output that will come from the collaboration...should then hold each other accountable to these expectations once they’re set by checking in about them at...speed up the future directions of the project. The state of research is constantly shifting, and long-distance... -
CUT&RUN: An Improved Method for Studying Protein-DNA Interactions
TypeBlog Post... does have limitations. The enzyme-based CUT&RUN method overcomes many of these limitations and makes ...graduate student at UNC Chapel Hill. Chromatin immunoprecipitation followed by high-throughput sequencing, ChIP-Seq... binding patterns that overcomes some of the limitations of ChIP-seq. CUT&RUN, which stands for ‘Cleavage... -
CASTing Off for New Shores in Human Genome Editing
TypeBlog Post...assistance). Quicker CAST activity allowed certain mutated phages to outcompete their brethren, and the researchers...Witte et al., 2025). The best evoCAST featured 20 mutations in TnsB, the subunit that catalyzes DNA insertion...therapy, highly tailored treatments to specific mutations require individualized optimizations, safety testing... the National Academy of Sciences of the United States of America, 114(35), E7358–E7366. https://doi.org... -
FastCloning: A Newer, Simpler Cloning Technique
TypeBlog Post...several struggles with these techniques: unwanted mutations due to excessive PCR cycles or low fidelity Taq...purify and sequence the plasmid to confirm no mutations were introduced during PCR or any subsequent steps...fidelity DNA polymerases, I have never had any mutations introduced into my inserts. However, with any ...technique there is a chance of introducing an unwanted mutation. Tips for high efficiency cloning Now that you...