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We narrowed to 762 results for: ire;

Showing: 261 - 280 of 762 results

  1. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...in different CGBE tools allow the researcher to direct the repair of the abasic site. But what if an editor...transversion editors. Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows indicate...this point relied on deaminase enzymes, either to directly accomplish the base edit or to create the substrate...they realized that MPG could be engineered to act directly on unmodified G bases, resulting in a G → Y (Y...Conclusion With these advances, every base can now be directly targeted by a base editor, although the efficiency...

  2. The Challenges of Cell Culture

    Type
    Blog Post
    Published
    May 5, 2016, 2:30 p.m.
    ...contamination: Epithelial Mesenchymal Transition Is Required for Acquisition of Anoikis Resistance and Metastatic...from the official biorepositories or have been directly isolated from patients. In these cases, a test...outlined in this article. Now many journals strictly require evidence of cell line provenance and identity. ...Council (BBSRC) and Medical Research Council (MRC) require authentic cells to be used and STR profiling to...can be prevented and we can progress faster to desired clinical endpoints.   Nick Amiss is an ATCC Outreach...

  3. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Published
    Nov. 5, 2014, 3:41 p.m.
    ...Historically, molecular interactions were detected by indirect means often using probes with the potential to...proteins, pioneers of this technique were able to directly detect protein/protein interactions in living ... to each other in the spectrum, you can easily directly excite the acceptor with the laser used to excite...corrective approaches and appropriate controls are required to make this method useful for dynamic experiments...fluorescence, this method is also less sensitive to direct acceptor excitation artifacts and it is possible...

  4. Transferable Skills Guide: Time Management

    Type
    Blog Post
    Published
    Dec. 5, 2017, 1:52 p.m.
    ...3 days to complete the entire experiment, but you’ll have to spend the entirety of each day doing the ...work in a job where you’re pulled in a thousand directions at the start of the day, writing your to-do list...depositors from Boston University Contact Dr. Gilmore directly to see if he knows anyone who’s interested in ...if I had planned my day extensively, my lack of desire to go sit in the tissue culture hood may have prevented...

  5. Five Popular Model Organisms

    Type
    Blog Post
    Published
    April 11, 2019, 12:39 p.m.
    ...interest (Fire et al., 1998). The discovery of RNAi in the worm actually rewarded depositors Andrew Fire and...Yeast are also the first eukaryotic genome to be entirely sequenced and is very amenable to genetic manipulation...utilized the worm to study calcium signaling in the entire adult brain, using a plasmid that expresses a genetically...(1974): 71-94. PubMed Central PMCID: PMC1213120. Fire, Andrew, et al. "Potent and specific genetic interference...

  6. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...instability at 37°C (Ouellet et al., 2016) and requires a tRNA scaffold to improve its stability in vivo...Ming Hammond’s group created Spinach2 using site-directed mutagenesis of Spinach. In doing so, they identified...green fluorescent aptamer, was identified through directed evolution. First, a pool of RNAs that activate...Fluorescence is observed when these RNAs complex and the entire aptamer can hybridize and fold. (Alam et al., 2017...fluorescent protein by fluorescence-based selection and directed evolution." Journal of the American Chemical Society...

  7. Plasmids 101: Biotinylation

    Type
    Blog Post
    Published
    Nov. 15, 2018, 1:50 p.m.
    ...Vitamin B7) is first an essential coenzyme that is required by all organisms for life. Biotin is only synthesized...Most organisms have less than five enzymes in the entire proteome that are biotinylated. In E.coli for example...responsible for biotinylating only one lysine in the entire organism on the biotin carboxyl carrier protein...recall, BirA biotinylates only one lysine in the entire E.coli proteome. Using this knowledge, scientists...scientists identified the 13 amino acid sequence required for BirA recognition and biotinylation. This 13 amino...

  8. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    Published
    Jan. 4, 2022, 2:15 p.m.
    ...a stupendous listener, but I aspire to  be. If you don’t at least aspire to be a good listener and consciously...each other disagree about a specific decision, direction, or goal. Resolving these types of conflicts is...Assumptions: Making incorrect assumptions can be a direct cause or consequence of lapsed communication. Making...job”. It’s certainly resolvable, but will likely require a manager to explicitly assign the responsibility...

  9. Antibodies 101: Normalization and Loading Controls for Western Blots

    Type
    Blog Post
    Published
    April 8, 2025, 1:15 p.m.
    ...sample would directly correlate to protein abundance on your membrane, which would be directly reported by...housekeeping proteins.” Fully quantifying a western blot requires a standard curve with a pure sample of your protein...proteins may not transfer precisely evenly across the entire blot. So how can you accurately compare different...your approach) as your final blot. Quantifying the entire lane of protein provides a loading control that...

  10. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    Published
    Dec. 2, 2019, 2 p.m.
    ..., BioID2 exhibits enhanced proximal labelling, requires less biotin, and allows scientists to more selectively...stability. Thus they conducted multiple rounds of directed evolution until they identified a mutant with ...APEX2, although it can rapidly label proteins, requires H2O2 which can be toxic to living cells. To circumvent...issues, the Ting lab utilized yeast display-based directed evolution to generate two mutants, the 35 kD TurboID... PMCID: PMC4831873. Lam, Stephanie S., et al. "Directed evolution of APEX2 for electron microscopy and...

  11. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    Published
    June 23, 2016, 2:30 p.m.
    ...system (OTS) for the incorporation of phosphoserine directly into recombinant proteins (5). Optimization of...of this system defined three components required for translation (Figure 1): A tRNA that recognizes a... The generation of pSer-incorporated proteins requires a molecular toolkit containing the OTS, an appropriate...phosphoserine OTS, Sep-OTS, contains the components required for the translation of pSer-incorporated proteins...phosphoserine must be recoded to ‘TGA’ using site-directed mutagenesis. In the C321.ΔA strain, this TGA will...

  12. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    Published
    June 1, 2023, 1:15 p.m.
    ...studies, where it could potentially label cells not directly connected to the injection site, or when using...volume. You’ll often hear that “high titer” is required for efficient AAV transduction, usually without...the optimal number of viral particles per volume required to express the gene of interest, in the cells ...are a few factors to consider. First, the titer required is inversely related to transduction efficacy:... recommended to optimize to the “lowest number required” instead of using the first amount that looks ...

  13. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...transformants for single colonies (this usually requires 10-20 plates) (see box below). This initial transformation...growth, scrape up, pool and extract plasmid DNA directly from the colonies. Transform the resultant plasmid...a lysis buffer to release recombinant proteins directly onto the nitrocellulose for subsequent probing... can lead to high background that obscures the desired signal. We have successfully corrected this by ...organism, it can be helpful to spot this protein directly onto nitrocellulose and carry out the steps from...

  14. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ...way to verify that the insert ended up in the direction you want - usually by Sanger sequencing or further...course there’s much more detail and verification required for the process to work well, so let’s go over... that the digest go until completion. The time required for complete digestion varies for different enzymes...associated manufacturers websites. These stains require you to either stain your gel after you run your...above, respectively). Some of the above stains require you to visualize your DNA using UV light – please...

  15. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    Published
    Nov. 28, 2017, 2 p.m.
    ... Material Transfer Agreement process that kits require; Nicole, the web master in charge of all our externally...You might find that your next paper is going to require help from a biochemist, a geneticist, a microscopist...world, so a process or project or mission that requires a broad knowledge base is going to necessarily...more things necessary to make a successful team. Entire books have been written about this! But this is...the type of team. Knowing what roles your team requires to function is certainly necessary, but equally...

  16. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    Published
    Oct. 17, 2019, 12:28 p.m.
    ...membrane, a threshold is reached and the neuron fires, sending the electrical signal down the axon (Lodish...subsequently photo-stimulated through fiberoptic cannulas directly implanted in the injection site and connected ... et al., 2008; Taylor et al., 2016; Hooper and Maguire, 2016). Recombinant adeno-associated viruses (rAAVs...Central PMCID: PMC4971574. Hooper, Andrew, and Jamie Maguire. "Characterization of a novel subtype of hippocampal...PubMed PMID: 4940442. Schnütgen, Frank, et al. "A directional strategy for monitoring Cre-mediated recombination...

  17. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ... two very similar proteins. Additionally, they require variable expression of your target across samples...and it causes a large deletion that removes the entire exon that codes for the epitope recognized by your...expected target protein in a Western blot. You acquire a plasmid encoding your protein of interest, known...you very similar results, while the third looks entirely different (B), giving you confidence that your...amenable to high-throughput analyses, however, it requires access to a mass spec facility and the ability...

  18. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...Transposition protein: This is the protein that’s required for the transposon sequence to move from one DNA... transcriptase and integrase enzymes which are required for LTR retrotransposon transposition. Non-LTR...animals Transgenic animals are often generated by directly injecting DNA into the pronuclei of fertilized...dystrophin, the gene mutated in muscular dystrophy, require truncation to fit in viral vectors, but easily ... into eukaryotic and prokaryotic genomes. They require just two key components, the transposon’s DNA sequence...

  19. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    Published
    Sept. 17, 2024, 1:15 p.m.
    ...concentration of your reagent. Be aware that this approach requires excellent attention to detail, since you’ll end...Transfers are a little trickier than gels and may require multiple runs to optimize or troubleshoot.   Before...migrate…so don’t try eliminating one component entirely. If you are trying to transfer proteins of vastly...antibody.  Pro tip! If your antibodies are coming directly from tissue or supernatant, and you’re doing the... you don’t need to compare the protein amounts directly, stripping and reprobing. Just make sure to go...optimizing it. Blots for quantification generally require more optimization than blots used to confirm overexpression...

  20. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    Published
    Oct. 24, 2017, 1:54 p.m.
    ...in a hyper-competitive environment where getting hired hinges on performing near miracles. When evidence...defending the role of science in our democracies will require constant vigilance and actions beyond marches, ...idea, but they might feel disconnected from the entire scientific process.  I've spent a lot of time debating...debating how to fix this.  As scientists, we work tirelessly in the lab where the groundbreaking research ...training as an academic scientist. I’ve worked tirelessly to get the publications, grant funding, and teaching...
Showing: 261 - 280 of 762 results