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  1. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...Ribosome display The Seeger lab starts with a “pre-enrichment” selection step using ribosome display because...size (up to 1012 members) in comparison to other display systems using phage and yeast (109-1010 members...ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated target ...can be recovered for the subsequent steps. Phage display mRNA from the enriched pool of sybody candidates...produce a phage library consisting of phages that display the candidate sybodies in their coat protein to...protocol describes going through two rounds of phage display to increase the enrichment of target-specific sybodies...ELISA) The enriched sybody pools from the phage display screens are subcloned (using FX cloning) into the...
  2. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ...“I’ll be there in a few minutes, I just need to split some cells.” Inevitably, this task would take longer...prioritizing. In my anecdote above, I prioritized splitting cells over meeting up with friends, but didn’t...terms of the utility of your tasks. In my cell splitting anecdote, I probably could have grabbed some food... food with friends and come back to the lab to split my cells later. Eating with friends has a somewhat...all the utility (fun) of eating with friends by splitting my cells first. Time management through planning... I probably wouldn’t have fallen into my cell splitting issue at all if I’d planned appropriately. Back...tissue culture hood may have prevented me from splitting cells until late in the day anway. There are a...
  3. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...miniturbo Split-BioID: A conditional proteomics approach to monitor protein-protein interactions Split-BioID... assemble when in close proximity. Split-BioID consists of a split BirA* molecule that can reassemble ...circumvent these issues, the Ting lab utilized yeast display-based directed evolution to generate two mutants...complex of interest. For more information, read the Split-BioID blog post.  RaPID: RNA-protein interaction...(LPETG). In EXCELL, the mgSrtA variant can be displayed on the surface of a cell of interest, where it...PMCID: PMC3916822. Schopp, Isabel Myriam, et al. "Split-BioID a conditional proteomics approach to monitor...
  4. CRISPR Guide

    Type
    Collection
    ... reverse transcription. The edited DNA strand displaces the unedited 5’ flap, and the resulting heteroduplex...including editors based on a second ADAR variant that displays improved specificity, and editors carrying an ... in vivo RNA imaging or to control alternative splicing. Figure 13: Overview of RNA targeting (A) and ...locus-specific targeting of long RNAs with CRISPR-Display. Nature Methods , 12 (7), 664–670. PMID: 26030444...
  5. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ...NEBridge SplitSet If you want to design an overhang set specific to your DNA sequence, SplitSet is the .... Once you input a sequence, SplitSet finds optimal fusion sites to split it into multiple fragments. ...minimum number of fragments, or defined split regions. SplitSet will then return the highest-fidelity overhang...Fidelity Viewer™; NEBridge GetSet™; and NEBridge SplitSet™. With these tools, NEB successfully performed...
  6. Rett Syndrome

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    Collection
    ...RNA editing, gene replacement therapy, RNA trans-splicing, and protein replacement to reverse the impacts...2002. Insight into Rett syndrome: MeCP2 levels display tissue- and cell-specific differences and correlate...
  7. Zhang Lab's CRISPR Frequently Asked Questions

    Type
    Collection
    ...because the top figure shows the construct from our 'Split RNA' design, where the pre-processing form of crRNA... more details. One important note is that this ‘split RNA’ design is not as efficient in genome editing...
  8. Plan Your Experiment

    Type
    Collection
    ...should be made HDR requires a repair template and displays reduced efficiency compared to NHEJ knockout. ...region is removed from the mRNA due to alternative splicing. Exons near the N-terminus are targeted since ...
  9. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    ...score, and type of article. Now, search results display a bar graph of the number of papers with your search...papers with your key term published each year is displayed with your search results and can be used to filter...other manuscripts that heavily cite the paper are displayed. You can also limit searches to particular authors...entry, the number of citations that paper has is displayed at the top of the page, as well as the number ...background, methods, or results section is also displayed. Saving papers Papers can be saved to a Semantic...shared online, or its altmetric score, is also displayed in your feed. By clicking on a paper in your feed...
  10. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...on this tool by splitting cGAL4 in two and binding each half to a gp41-1-N-intein. Split inteins are protein...PMC5693259.  Wang, Han, et al. "Split cGAL, an intersectional strategy using a split intein for refined spatiotemporal...knock-down proteins of interest.  Plasmids for cGAL and split cGAL systems The Sternberg lab in 2016 created cGAL...peptides together (Wang et al., 2018).” With the split system, only cells that have both halves of cGAL...
  11. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...contaminants. Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine..., for example, split cells on a Monday diluting them 1:10, you should be able to split on the following...Thursday, or, at the latest on Friday. It’s best to split cells before the plates are more than 80-85% confluent...metabolism that is toxic to the cells), and need to be split very soon. There is power in numbers! When starting...
  12. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...reporters altogether; In the split-luciferase system, intein-mediated splicing reconstitutes functional luciferase... luciferase protein interact and enable intein splicing (5). Screening with yeast two hybrid systems While... this requirement for transmembrane proteins, a split ubiquitin system has been devised (8). References...Kaihara, A., Sato, M., Tachihara, K., Umezawa, Y.l. Split luciferase as an optical probe for detecting protein-protein...interactions in mammalian cells based on protein splicing. Anal Chem. 2001; 73:2516-2521. Pubmed PMID: 11403293...
  13. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    ...reporter, ZipGFP, which utilized the self-assembling split GFP (GFP1-10 and GFP11) to increase fluorescence... this they modified a different self-assembling split GFP (GFP1-9 and GFP10-11) by flipping a single β-strand...11 β-strands and one central ɑ-helix that can be split into 3 parts: (1) β-strands 1-9 (β1-9) and the one...superfolder Cherry (sfCherry). Like FlipGFP, sfCherry was split into  β1-9 and β10-11, where β10-11 was forced into...-protein interaction sensor based on tripartite split-GFP association." Scientific reports 3 (2013): 2854...
  14. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ... some you’ve likely never heard of. Each entry displays the following information for a Cas enzyme: ...properties Links to relevant resources The homepage displays a graphic describing CasIDs and relevant properties...feature. Activity features: CasID CasPEDIA will display both the cis and trans activity of each entry. ...protospacer-flanking sequence (PFS) in some cases. The resource displays these requirements in regard to their sequence... covered!     Fig. 1 – CasPEDIA Cas ID display information for a given Cas (SpyCas9a) from CasPEDIA...
  15. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...excited to announce that our plasmid and sequence displays are now powered by GSL Biotech's SnapGene Server...feature library, our updated plasmid and sequence displays are now much easier to interpret and analyze at...CRISPR gRNA cloning. With this more functional display, you can start thinking about your next cloning...experiment early on. Beyond these simple but powerful display improvements, when you click on any plasmid map...enhancements that will further improve the map displays and functions. ...
  16. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...scientists can mutate the transcription start site, splice sites, exons, and/or promoters to knock out lncRNA... and Bester 2021; Zibitt et al. 2021). Mutating splice sites is particularly effective as it can result...Transcription start site (TSS) deletion. (B) Mutations of splice sites. (C) Removal of an exon or a large genomic...noncoding RNAs in human cells by Cas9 targeting of splice sites. Nat Biotechnol. Published online November...expression by lncRNA promoters, transcription, and splicing. Nature. 2016;539(7629):452. doi:10.1038/nature20149...
  17. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    .... This style displays a poster vertically rather than horizontally. Instead of displaying your information...employing a large central box, often used for displaying all of one’s figures, or at least the majority...inserted. The advantage of this style is that it will display your data most prominently, making it the star ...information as columns, you may choose to display it as rows. It can be more catchy because it is generally...
  18. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...range of PAMs like NG, GAA, and GAT, but also displaying increased editing specificity. We’re excited ...recognizing NG, NNG, GAT, and CAA PAMs, while xCas9 3.6 displayed the second best PAM range. Characterizing xCas9... SpCas9 in mammalian cells, finding xCas9 3.7 displayed about the same editing rate as SpCas9 at an NGG...SpCas9. For some non-promiscuous gRNAs, xCas9 3.7 displayed more than a 100 fold decrease in off-target editing...ratio. Importantly, the observed off-target loci displayed a broad range of PAM sequences, indicating they...
  19. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...detecting protein-protein interactions that uses split fluorescent proteins tethered to potential protein...visualize PPIs. This fluorescent protein (FP) is split into two fragments that individually lack fluorescence...fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent protein, YFP) is reunited...information about the structures of the tested PIPs. Many split FPs have been validated for BiFC (Kodama and Hu,... Hu, 2012). Such controls are necessary because split FPs can associate, albeit at relatively low frequencies...
Showing: 21 - 40 of 230 results