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We narrowed to 727 results for: cat.1

Showing: 361 - 390 of 727 results

  1. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ...your budget, you should take the following steps: 1. Define your timeline Start with your thesis in mind...these decisions in the following order of importance: 1. First make decisions based on experimental timelines... several factors aren’t taken into consideration. 1. Always focus on the science When negotiating finances.... After identifying all the major equipment, categorize all of your minor tools, like pipets, or mini-centrifuges...titles in the regions where the host institute is located. For example, salaries in Kansas versus salaries... most start-up P.I.’s. It doesn’t have to be complicated, or difficult to create this list. Start BEFORE...discounts, collaborations, access to internal award applications, etc. If this is the institute that can help...

  2. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    .../nmeth.2641. Epub 2013 Sep 1. PubMed. Find plasmids from this publication at Addgene. Scalable and...sequences are inserted into the genome. Figure 1: Self-excising cassette schematic from Dickinson et...genomic modifications, is very robust in C. elegans, and the methods for HDR-mediated modification continue...After self-excision, the resulting loxP scar is located in this synthetic intron, and only the fluorescent...use and highly efficient for C. elegans genome modification. Figure 2: Schematic from Dickinson et...permission from the Genetics Society of America. Applications of this system Dickinson et al. have developed...system could be used to make many other genome modifications, including targeted mutations. SEC-based systems...

  3. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    ...those ideas and build from those.   Figure 1: Virtual lab meeting? No problem! You can still creatively... on it – so it is important to take breaks. After 1 hour (or another designated time) of meeting, someone...Collaboration and creativity are pivotal in writing publications/grants too. As described by several works (Wuchty...backgrounds.” This work outlines the need for clear communication, fair credit through authorship, and acceptance...remotely. The authors stress the fact that “miscommunication, a lack of leadership, and inappropriate tools...approaches can lead to frustration, delay of publication, or even the termination of a project.” Take ...setting. Check out the blog post “Fostering Communication in Remote Collaborations” to get more ideas....

  4. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ... are not required to initiate the reaction. Table 1 compares some general advantages and disadvantages...disadvantages of luminescence versus fluorescence.  Table 1: Properties of Luminescence versus Fluorescence   ...Luciferases are a class of enzymes capable of catalyzing chemical reactions in living organisms resulting...gene, such as luciferase, usually serves as an indicator of transcription within cells, where detection...photon emission is measured resulting from the catalysis of a chemical reaction requiring luciferin, ATP...Background Lower Higher Post-translational Modification Not Required Required Photobleaching/Phototoxicity...blue light with a peak emission at 480nm upon catalyzing a chemical reaction with the substrate coelenterazine...

  5. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...sequences were initially discovered in E. coli in 1987 (1), the concept that these clustered repeat sequences...neutralize CRISPR/Cas. Earlier this year, expression of 1 of 5 anti-CRISPR genes were found to inactivate the...distinct types and subtypes using polythetic classification based on evolutionary similarity (5). Type ...2013 (7, 8). Nine months later, more than 1500 publications describe work to improve the tool’s specificity...various species and describe an assortment of applications. Despite the ubiquity of CRISPR/Cas toolkit,... multiple plasmids used for various platform applications. For more information on CRISPR/Cas, check out...for your research. Do you have a cool idea or application that involves CRISPR/Cas technology? Let us know...

  7. Data Freedom: The Expansion of Data Sharing in Research Publications

    Type
    Blog Post
    ...processed since the declaration, only a small fraction (<1%) of authors have asked for advice about the scope...where readers may locate data supporting the research reported in PLOS publications. The policy was not... is founded on the principle that scientific publications must contain enough information (or links to...probable to some extent - but that is the price of publication. Who is to say that the initial data-generating...logically annotated and arranged, as in an actual publication and not require interaction with the data authors...likely, at least in part, due to our inadequate cataloguing of how results were obtained and this alone is... Nature Publishing Group recently launched a publication called Scientific Data, which aims to act as ...

  8. Chemogenetics Guide

    Type
    Guide
    ...out our Chemogenetics Plasmid Collection . Figure 1: General overview of a chemogenetics experiment. Early...bidirectional control of neuronal activity. Table 1. Descriptions of DREADDs and their activity in neurons... while PSAMs and LMOs have an effect for only 0.5–1 hours after delivery. Chemogenetic ligand. The chemogenetic...neurons) that can be turned on or off by the application of a small molecule ligand. The ideal chemogenetic...muscarinic Arrestin-2/-3 CNO* Increase Arrestin translocation Arrestin signalling Nakajima & Wess, 2012 hM3Dq...chemogenetics for research and potential clinical applications. Science. 364(6436):eaav5282. PMID: 30872534... KM, Brown KM, Forbes SL, Liggett SB (2001). Modification of the beta 2-adrenergic receptor to engineer...

  9. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    ...distinguish between your two targets.    Figure 1: In the isotype conversion process the variable regions...natural isotype to that of a different group (Fig. 1). This process, called isotope conversion, gives users...published, knockout validated, and work in your application. But…they are the same isotype and therefore ...are identical for antibodies within certain classification groups called isotypes.  Why does isotype matter...

  10. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    ...restriction digests are comprised of 2 separate steps: 1) incubating your DNA with restriction enzymes which...The resulting gel image includes a 1kb ladder (lane 1) that has bands ranging from about 500bp to 10kb, ... I hope these tips demonstrate that plasmid verification is not just necessary but also an easy process.... Please vist Addgene's resource for plasmid verification to find additional tips and detailed protocols...

  11. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    ...activity degrades the displaced strand.   Figure 1: EvolvR consists of an error-prone polymerase (PolI...mutation rate and extend the editing window of EvolvR: (1) two additional mutations to PolI3M generated PolI5M...potential applications. Challenges of existing mutagenesis methods Several continuous diversification methods... challenging for users to achieve targeted diversification of all nucleotides within a defined region ...Chang Lab have created methods that introduce diversification by targeting a site of interest with dCas9 ...level of variation that can be introduced.   Diversification is limited due to the integration of oligonucleotide...increasing the standard mutation rate seen during DNA replication by 120-fold over background. Due to its modular...

  12. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    ...sticks out and can be considered a linker (figure 1). In fact, for several FRET biosensors (yellow cameleon...fluorescent protein are relatively close together (figure 1). Where should the fluorescent protein be inserted...fluorescent protein database." Nature methods (2019): 1. PubMed PMID: 30886412. Mastop, Marieke, et al. "A...study its function is one of the most popular applications of fluorescent proteins. These fusion proteins...steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding sites or motifs that...that are necessary for post-translational modifications. Despite these potential issues, GFP has been used...linker sequence short In the early days of GFP applications, many were concerned with steric hindrance by...

  13. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...overcomes many of those limitations (Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule...GTPase Signaling Toolbox. ACS Synthetic Biology, 11(1), 515–521. https://doi.org/10.1021/acssynbio.1c00604...efficiency constructs are great for in vitro biocatalytic applications where toxicity doesn’t matter. Plus, for... binding of the plasma membrane. In previous publications, they described tools to control formation of.... Restricting the effect to a specific time or location can be critical when, say, the gene is essential...that trap the fusion protein away from its usual location, preventing it from carrying out its proper function...) and to introduce unnatural head groups for applications like fluorescent labeling via click chemistry...

  14. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...engineered human uracil DNA glycosylase. Nature Communications, 15(1), 4897. https://doi.org/10.1038/s41467-024...engineered uracil-DNA glycosylase. Nature Communications, 15(1), 6397. https://doi.org/10.1038/s41467-024...a glossary at the end of this post! Figure 1: Base transitions are edits between the purine bases...Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows indicate natural cellular...researchers can now select the best tool for their application from among many options.  Conclusion With these...

  15. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...bacteria with superior performance. Nature Communications, 9(1). https://doi.org/10.1038/s41467-018-04899...oxygen. Additionally, its status as a Biosafety Level 1 microbe makes it a perfect candidate for introductory...Transport, biofilms, and much more. Because of it’s BSL-1 status, E. coli can be used as a model to relate these...studying genetic modification for their ease of manipulation and their ability to be replicated. Because E...., 2004). Additionally, breakthroughs in gene modification tools and methods such as CRISPR have vastly...Escherichia coli as a Model Organism and Its Application in Biotechnology. In Escherichia coli - Recent...Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306 Koornneef...

  16. Evolution of Lab Techniques

    Type
    Blog Post
    ...blotting, named for its inventor Edward M. Southern (1), can be used to detect DNA identity, size, and abundance...hours at low cost with restriction mapping (Figure 1). Though some labs may prefer to send DNA directly... serotonergic system and behavior.     References 1. Southern, Edwin Mellor. "Detection of specific sequences...comparison to a control sample. Southern blotting applications range from confirming the results of cloning...interest by recording DNA abundance throughout the amplification process, rather than just at the end as in standard...experiments allowed researchers to identify the location of genes. With this knowledge, researchers were...was added and when. Compiling the identity and location of a given nucleotide, a researcher (or a simple...

  17. Introducing the Addgene Antibodies 101 eBook!

    Type
    Blog Post
    ...antibodies as research tools, you may find “Chapter 1: What is an Antibody” quite useful. If you’re looking...resource for anyone looking to use antibody-based applications in their research.  Like all our eBooks, the...through the process. If you’re confident in your application and protocols, but can’t quite seem to find an...for Your Experiment.” Whatever your antibody application question, the Antibody 101 eBook is a great place...

  18. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    ... There are two methods of neuronal tracing (Fig. 1): one to visualize neurons that send information to...significantly updated in 2023 by Jason Nasse.  References 1. Tervo, D. Gowanlock R., et al. "A designer AAV variant...Tracers are typically delivered to a particular location and they either trace downstream from the soma...Others have noted a lack of transduction in catecholamine pathways from the spinal cord (Wang, 2018, Metcalfe...not yet understood, one thing is clear: this application opens a lot of doors in neuroscience. The retrograde...retrograde serotype also has promising therapeutic applications; it can enhance therapeutic delivery across ...

  19. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    ... to be aware of the April 1 deadline for companies to submit an application on their behalf.  If an individual...and non-profit organizations do not have the April 1 H-1B queue process or quota like industry, these organizations... National Interest Waiver. The three prongs are: (1) The foreign national’s proposed work endeavor must...-1B, you need to prove 3 threshold requirements: (1) that you have international recognition for your ...higher education nonprofit entities related to or affiliated with institutions of higher education nonprofit...for self-sponsorship of the I-140 application meaning that the category does not require a foreign national.... NIW requirements stress “educational background, expertise, publications, and plan for future activities...

  20. Predicting Adverse Reactions to Monoclonal Antibody Drugs

    Type
    Blog Post
    ...immunogenicity score (Mattei et.al., 2022).    Figure 1: Generation of immunogenicity scores by predictive...antibodies for the treatment of diseases. J Biomed Sci 27, 1 (2020). https://doi.org/10.1186/s12929-019-0592-z  ...silico approach The in silico approach cannot replicate the complexity of antigen processing events that...This system would somewhat act like a 2-step verification. All approaches have their own limitations. ... together can inform decisions about in vivo application and clinical testing. This would pave the way...

  21. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...restriction digest and colony PCR.   Fig. 1: (A) Schematic of cloning, expression and validation... doi: 10.1371/journal.pone.0038313. Epub 2012 Jun 1. PMID: 22675541; PMCID: PMC3365890....cDNA, which  was used as a template for PCR amplification with a degenerate primer set to amplify IgG ...  While screening by IF-ICC worked well for identification of the correct clones, it was determined that...through functional assays. Following functional verification, the R-mAb expression plasmids were DNA sequenced...

  22. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...done.   Figure 1: DNA melting by DnaA and IHF is the first step of replication for the bacterial chromosome...plasmids rely on their host cell's replication machinery in order to replicate—but not always to the same extent...described in our previous Origin of Replication post, DNA replication is initiated at the ori and may or...consider the replication mechanism of plasmids under stringent control, such as pSC101. DNA replication and regulation...review how E. coli normally replicate their chromosomal DNA. Chromosomal replication in bacteria is carefully...entire round of replication. Initiation is the primary control point of DNA replication in bacteria, so... the replication forks reach each other on the opposite end of the chromosome, ending replication.   ...

  23. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...first foray into Reddit. Here are some reasons why: 1. Reddit has a large and active community of people...re more interested in selling a product than communicating your ideas with people or discussing science...to write about in the future. The AMA was also cataloged for permanent reference by the Winnower. The only...I do a lot of public speaking where I enjoy communicating face to face with the audience. I post on LinkedIn...own expertise and your resources for science communication opportunities like an AMA. One of the best things... science communication...

  24. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...modified slightly with either a 1-7 base poly(U)tail in tRNA-gRNA constructs or a 1-4 base tail from the second...biological pathways;  to observe a phenotype, the modification of multiple genes is often necessary. Guide ...recognition and cleavage. This sequence flexibility indicates that gRNAs could be incorporated into these constructs...the 5’ end. PTG construct schematics. Scissors indicate cleavage sites. After confirming gRNA cleavage...) How many loci can Cas9 modify? To test the application of this system to gRNA multiplexing, Xie et al...deletion with 4-20% frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct...multiplexing further illustrates the potential of PTGs. Applications of PTGs The major advantage of the PTG system...

  25. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    ... and, if you do it right, can be (part of) figure 1 of your next paper or thesis. This series of posts...An explanation of the assay is depicted in figure 1. The strict correlation between the fluorescent protein...protein for your application. Selecting the best fluorescent protein for a specific application can seem like...under conditions that best mimic the intended application. The three key properties for live cell imaging...are converted into emitted photons. These are indicated by the extinction coefficient (EC) and quantum...experimental set up. The practical brightness takes all application specific parameters into account, including the... picture of what can be expected in a 'real' application than the theoretical brightness. Consequently...

  26. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    ...schematic depiction below): 1. Resistant cells among the separate replicates would contain the same labels...development of combination therapies.         References 1. Hyo-eun, C. Bhang, et al. "Studying clonal dynamics...summarized in a recent Nature Medicine paper. In the publication, Bhang and coworkers used the ClonTracer library...propagated it, separated it into roughly identical replicate groups, and treated these groups with the same...subpopulation 2. Resistant cells among the separate replicates would have different labels - i.e. they would... Looking at barcode abundances among multiple replicate populations, they found that similar barcodes ...populations were shared by at least one other replicate; prior to treatment, there are small (previously...

  27. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...Introduction to Biosensors. Essays Biochem. 2016 Jun 30;60(1):1-8. DOI: 10.1042/EBC20150001 Leopold, A. V., Shcherbakova...of the biosensors used in research are very sophisticated, but at their core they can all be boiled down...ready-to-use. While biosensors span the gamut of applications within chemistry, biology, and physics, in this...sensors can be targeted to different cellular locations or tissues depending on the biology you are studying...different factors, in different or the same cellular location, all at once! Small molecules What type of small...Neurotransmission and Neuromodulation: Engineering and Applications. Frontiers in Cellular Neuroscience. 2019 Oct...

  28. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    ...protein of interest will be problem-free. In figure 1 we provide an example of the OSER assay with a number...can be shared by publishing it as (part of) figure 1, uploading it as a preprint, or documenting it in ...aggregates. This property can be tolerated in some applications (e.g. labeling of cells or tracking promotor...promotor activity), but it is problematic in applications in which the fluorescent protein is used as an inert...During the experiment, the solution is diluted, complicating the interpretation. Because of this, these methods...aware that the fluorescent proteins reported in publications are only characterized under a limited number...build a valuable resource with tools and data that indicate the performance of (a set of) fluorescent proteins...

  29. Suggest A Plasmid You'd Like to Find at Addgene

    Type
    Blog Post
    ...to reach out to potential depositors in two ways: 1. Fill out our "Suggest a Plasmid" form Fill out the...suggestaplasmid, the name of the plasmid and a publication where we can find the plasmid. While we cannot...

  30. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ...bioengineering, but it wasn’t clear whether they could 1) be decorated with proteins and remain functional,...Follow her on Twitter @jessicapolka.     References 1. Kanabrocki, J. A., Quackenbush, R. L. & Pond, F. ... modified to extend at different pH levels. Domesticating R bodies We have shown that all three of these...actuators that could be used in a variety of applications. For example, you could use them to lyse cells...matrices) in response to pH changes. Beyond these applications, purified R bodies could also perform as actuators...PubMed PMID: 26814170. Find Plasmids from This Publication at Addgene 4. Petroni, G., Spring, S., Schleifer...
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