...and disease states, their circadian regulation, as well as in important physiological states such as adipogenesis...which then return information such as nutritional status. Within the animal kingdom, this information is...that interact directly with NRs) exist in steady-state complexes with secondary coregulator (or co-coregulator...whole complex.  For example, the phosphorylation status of SRC-3 defines its association with other members...

... PE6e-g editors with mutations to the Cas9 domain but found that these mutations only improved efficiency...created and evaluated 19 PE1 variants with RT mutations known to increase activity, enhance binding between...additional nuclear localization sequences; and two mutations in Cas9 previously shown to improve nuclease activity...effective prime editor proteins, but found that the mutations that emerged depended strongly on the target edit... ~ ~ ~ ✓ ✓ ✓ R221K N394K Cas9 mutations Improve Cas9 nuclease activity ~ ~ ~ ...

...antigens not present on the cell surface. The presentation of antigens can also be affected by cell fixation...quantified using the Attune NxT Flow Cytometer with representative images showing the staining intensity in the... Correctly interpreting your data therefore necessitates careful design of RNAi sequences, confirmation...validation is critical to ensuring accurate interpretation of your results. Therefore, taking advantage...Uniprot ID P21579) for use in western blot, immunoprecipitation, immunofluorescence and flow cytometry. F1000Research..., Robertson, J., & McPherson, P. S. (2019). Implementation of an antibody characterization procedure and...

...components of the plasmids to partial, full, or annotated plasmid sequences. These sequences can also be...the deposited sequence(s) in SnapGene. SnapGene annotates the plasmid maps based on SnapGene’s “Common Feature... function of the plasmid. This includes point mutations, frame shifts, missing elements, and nucleotide...plasmid that are less likely to affect function when mutated; elements that are difficult to sequence; approaches...Sequence discrepancies are not always a result of mutations in the plasmid, but can arise due to errors in...

...poster, you shouldn’t hesitate to be creative. After all, your poster is a representation of your work and ...most prominently, making it the star of your presentation. Portrait. This style displays a poster vertically...Morrison, a psychology doctoral student at Michigan State University. The idea is that the main research finding... center of the poster in a large font. Hence, spectators can read the text from many feet away and immediately...

...range of motion, a commutator allows the patch cable to connect to the laser and rotate 360° without a loss...wavelength to stimulate your opsin. Each opsin has an excitation spectrum (use the resources discussed in post...variant you wish to use to determine the ideal excitation). Lasers can be purchased through Laserglow, ...channel, Channelrhodopsin 2 (ChR2), exists in several states: closed/light receptive, open, closed/light insensitive...

...effects this mutation has on locomotion in our computer model to the effects of this mutation in the real...knowledge sharing, in the form of workshops, presentations, and lectures. There are currently around 1,...to modify the worm genome and make additional mutations in our worms.  We are on the lookout for genome...contributes each month - with only occasional supplementation with grant funds, money is always tight. Of...

...from New England Biolabs. Golden gate assembly limitations Embraced by the synthetic biology community, ...or two-step assemblies. We have reduced these limitations through the use of engineered Type IIS restriction...shown above) are flanked by BsaI sites in an orientation that results in 5’ four base overhangs after ...transformants onto the selection plate. Figure 2 shows representative transformation plates obtained from 1-, 12-...standard reaction conditions. This data set allows quantitation of sequence-dependent ligation efficiency and...

...Addgene makes this easy by providing a template citation for you to use in your Materials and Methods and...Addgene catalog number to easily find template citations for Addgene plasmids you have used in your research...depositor has indicated that there is a particular mutation found within the gene/insert, this information...questions about our plasmid pages, please don't hesitate to contact us at [email protected] Resources on... on the Addgene Blog Read about Our New Citations Feature Check out Our Walkthrough of the Deposit Process...

...expression? Are you trying to create a single point mutation? Do you want to create a fusion with a reporter...generating knockouts or knock-ins, or introducing mutations and tags, while a catalytically dead dCas9 can... Base editors can help you make precise point mutations near the gRNA target site. Choose an appropriate...repair (HDR) template to create knock-ins, point mutations, or to add a tag, you will also need to deliver... the expression system you choose in many ways dictates the best method for introducing those CRISPR components...

...chemical and peptide-based proteosome inhibitors, glutathione S-transferase (GST), which can be fused to recombinant...recombinant proteins for one-step purification with glutathione, can also protect against proteolysis. That’s... then your first step towards detecting, immunoprecipitating, or purifying that protein may be to fuse...Purification and Stability Good for purification with glutathione; protects against proteolysis, but may reduce...column runs. The TEV protease itself, with various mutations used to increase its stability activity, can be...

...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...

...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...

...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...

... 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...

...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...