...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...

...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...

...population and track connections to that population. Limitations of common genetically encoded neuronal tracers...attached to a protein of interest (POI), that facilitate immunohistochemistry (IHC) experiments with tag-specific... A new and improved neuronal tracer With the limitations of epitope tags and fluorescent proteins, it ...track with these classic probes. To overcome the limitations of existing FP and epitope tags, researchers ...Bobby Henderson, a physics graduate from Oregon State University. Find the Spaghetti Monster probes at...

...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...

...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. ...

...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...

...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...

...devised means of co-opting phage genetic systems to facilitate homologous recombination in bacteria. These quirks...non-edited cells to survive have inactivating mutations in the Cas9 or spacer sequence, and these rare...pneumoniae and can be used to generate multiple mutations simultaneously (Jiang et al. 2013). Genome editing...recombineering, with 96-99% efficiency for point mutations and 66% efficiency for small insertions. Multiplexing... E. coli, the system was used successfully in Tatumella citrea, another species of Enterobacteriaceae,...they were effectively able to introduce point mutations in P. aeruginosa, Pseudomonas putida, Pseudomonas...

...diversity in fluorescence brightness, intensity, excitation and emission spectra. The lab of Dimitri Deheyn...group to perform independent two-color optical excitation of neurons. The paper describes five previously...variety of contexts. In order to overcome these limitations, they evolved and characterized an improved peroxidase...Clonetegration could become a “valuable technique facilitating genetic engineering with difficult-to-clone ... sites that flank a DNA module in an inverse orientation, so that multiple DNA components can directionally...synaptic partners) technique based on functional complementation between two non-fluorescent split GFP fragments...combinations (depending on the number of copies). This facilitates the distinction of neighboring cells in imaging...

...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...

...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....

...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 ...

...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...

...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...

...elements often determines the type of plasmid and dictates how it might be used in the lab. Below are some...common plasmid types: Cloning Plasmids - Used to facilitate the cloning of DNA fragments. Cloning vectors... gene function as well as identify changes or mutations that may cause certain diseases. Sequencing DNA...

...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 ...

...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 ...

... 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...

...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...

...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...